Substituted pyrimidines and methods of use

ABSTRACT

The present disclosure provides inhibitors of activin receptor-like kinase 5 (ALK5). Also disclosed are methods to modulate the activity of ALK5 and methods of treatment of disorders mediated by ALK5.

CROSS-REFERENCE

This application is a continuation application of U.S. Ser. No.17/301,242, filed on Mar. 30, 2021, which claims the benefit of U.S.Provisional Application No. 63/002,658, filed Mar. 31, 2020; and U.S.Provisional Application No. 63/200,310, filed Mar. 1, 2021, each ofwhich is incorporated herein by reference in its entirety.

BACKGROUND

Human fibrotic diseases such as systemic sclerosis (SSc),sclerodermatous graft vs. host disease, nephrogenic system fibrosis, andradiation-induced fibrosis, as well as cardiac, pulmonary, skin, liver,bladder and kidney fibrosis, constitute a major health problem. Thesediseases often progress to organ dysfunction with eventual organ failureand death due to lack of treatment available, mainly because theetiologic mechanisms of runaway fibrosis are complex, heterogeneous, anddifficult to decipher. Activated myofibroblasts may be responsible forreplacing normal tissues with nonfunctional fibrotic tissue. Therefore,signaling pathways responsible for stimulating profibrotic reactions inmyofibroblasts have potential as targets for development of therapies totreat fibrotic diseases.

Normal tissue repair involves fibrotic reactions through homeostaticregulatory mechanisms. Uncontrolled fibrosis, however, may result inexcess deposition of the extracellular matrix (ECM) macromolecules ininterstitial space that stiffens over time. There are many sites alongthe molecular pathway leading up to myofibroblast activation, including,but not limited to, transforming growth factor-β (TGF-β) and bonemorphogenic protein (BMP) signaling pathways. Of importance in thisdisclosure is the pathway involving transforming growth factor-β(TGF-β), TGF-β receptor I (TGF-βRI), and TGF-β receptor II (TGF-βRII).

TGF-β signaling is typically initiated by binding of a TGF-β ligand to aTGF-βRII. This in turn may recruit and phosphorylate TGF-βRI, also knownas the activin receptor-like kinase 5 (ALK5). Once phosphorylated, ALK5typically adopts an active conformation and is free to associate withand phosphorylate Smad2 or Smad3. Once phosphorylated, Smad 2 and 3proteins then may form heterodimeric complexes with Smad4 which cantranslocate across the nuclear membrane and modulate Smad-mediated geneexpression, including, for example, the production of collagen. Smadproteins are intracellular regulators of transcription and therefore mayserve as modulators of TGF-β-regulated genes involving, inter alia, cellcycle arrest in epithelial and hematopoietic cells, control ofmesenchymal cell proliferation and differentiation, wound healing,extracellular matrix production, immunosuppression and carcinogenesis.

ALK5 is believed to be the most relevant of the activin-like kinases(ALKs) in the fibrotic process (Rosenbloom, et al., Fibrosis: Methodsand Protocols, Methods in Molecular Biology, 2017, Vol. 1627, Chapter 1,pp. 1-21). Several small molecules have been developed to inhibit theactivity of ALK5 for various therapeutic indications, related mostly tooncology (see Yingling, et al., Nature Reviews: Drug Discovery, December2004, Vol. 3, pp. 1011-1022).

SUMMARY OF THE INVENTION

One of the main problems with ALK5 inhibitors developed to date is thatthese molecules have been associated with ventricular or cardiacremodeling in preclinical safety studies resulting from significantsystemic exposure from oral administration. In view of the foregoing, aneed exists for small molecules that target ALK5 and for use of suchcompounds in the treatment of various diseases, such as cancer andfibrosis, while limiting adverse side effects. The present disclosureprovides these and other related advantages. One objective of thepresent disclosure is to deliver a potent ALK5 inhibitor locally withminimal systemic exposure in order to address any unintended andunwanted systemic side effects of ALK5 inhibition during treatment.Therefore, in some aspects, the present disclosure provides inhaled,long-acting and lung-selective ALK5 inhibitors for the treatment ofidiopathic pulmonary fibrosis. Compounds of the present disclosure maybe used to treat other diseases, including, but not limited to,pulmonary fibrosis, liver fibrosis, renal glomerulosclerosis, andcancer. Compounds of the present disclosure may be used as a monotherapyor co-dosed with other therapies, whether delivered by inhalation,orally, intravenously, subcutaneously, or topically.

In certain aspects, the present disclosure provides a compound ofFormula (I):

or a pharmaceutically acceptable salt thereof, wherein:

-   -   A and B are each independently selected from C₃₋₁₂ carbocycle        and 3- to 12-membered heterocycle;    -   R^(A) is independently selected at each occurrence from:        -   halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³, —S(═O)R¹,            —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,            —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —C(O)OR¹,            —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹,            —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂,            —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S, ═N(R¹);        -   C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, and —N(R¹)—C₁₋₁₀            alkyl, each of which is independently optionally substituted            at each occurrence with one or more substituents selected            from halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³,            —S(═O)R¹, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³,            —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹,            —C(O)OR¹, —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³,            —NR¹C(O)R¹, —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³,            —C(O)N(R¹)₂, —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S,            ═N(R¹), C₃₋₁₂ carbocycle, and 3- to 12-membered heterocycle;            and        -   C₃₋₁₂ carbocycle and 3- to 12-membered heterocycle,        -   wherein each C₃₋₁₂ carbocycle and 3- to 12-membered            heterocycle in R^(A) is independently optionally substituted            with one or more substituents selected from halogen, —NO₂,            —CN, —OR¹, —SR¹, —CH₂N(R¹)₂, —N(R¹)₂, —NR²R³, —S(═O)R¹,            —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,            —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —CH₂C(O)OR¹,            —C(O)OR¹, —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³,            —NR¹C(O)R¹, —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³,            —C(O)N(R¹)₂, —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S,            ═N(R¹), R¹, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, and            C₂₋₆ alkynyl;    -   R^(B) is independently selected at each occurrence from halogen,        —CN, —NH₂, —NHCH₃, —NHCH₂CH₃, —C(O)CH₃, —OH, —OCH₃, —OCH₂CH₃,        —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂F, —CHF₂, —CF₃, C₃₋₄        carbocycle, and 3- to 4-membered heterocycle;    -   m and n are each independently an integer from 0 to 3;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, 1- to 6-membered        heteroalkyl, C₀₋₃ alkyl-(C₃₋₁₂ carbocycle), and C₀₋₃ alkyl-(3-        to 12-membered heterocycle), each of which is optionally        substituted by one or more substituents selected from halogen,        —CN, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —CH₂CH₂N(CH₃)₂,        —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃,        —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₁₂ carbocycle,        and 3- to 6-membered heterocycle; and    -   R² and R³ are taken together with the nitrogen atom to which        they are attached to form a heterocycle, optionally substituted        with one or more R¹.

The compound of Formula (I) may be a compound of Formula (I-A):

For a compound of Formula (I) or (I-A), A may be selected from C₆₋₁₀aryl and 5- to 10-membered heteroaryl. In some embodiments, A isselected from phenyl and 5- to 6-membered heteroaryl. In someembodiments, A is selected from phenyl, indanyl, thiazolyl, thiophenyl,pyrazolyl, pyridyl, pyrimidinyl, indazolyl, benzotriazolyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzodioxanyl, andtetrahydrobenzazepinyl, such as A is phenyl. In some embodiments, A isthiazolyl. In some embodiments, A is thiophenyl. In some embodiments, Ais pyridyl.

The compound of Formula (I) may be a compound of Formula (I-B):

The compound of Formula (I) may be a compound of Formula (I-C):

For a compound of Formula (I), (I-A), (I-B) or (I-C), R^(A) mayindependently be selected at each occurrence from:

-   -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³,        —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹,        —NRIC(O)N(R¹)₂, —C(O)N(R¹)₂, —C(O)NR²R³;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is        independently optionally substituted at each occurrence with one        or more substituents selected from halogen, —OR¹, —N(R¹)₂,        —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂,        —C(O)N(R¹)₂, —C(O)NR²R³, C₃₋₈ carbocycle, and 3- to 10-membered        heterocycle; and    -   C₃₋₈ carbocycle and 3- to 10-membered heterocycle,    -   wherein each C₃₋₈ carbocycle and 3- to 10-membered heterocycle        in R^(A) is independently optionally substituted with one or        more substituents selected from halogen, —OR¹, —CH₂N(R¹)₂,        —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —NR¹C(O)R¹,        —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂, —C(O)NR²R³, R¹, and C₁₋₆ alkyl.

In some embodiments, for a compound of Formula (I), (I-A), (I-B) or(I-C), R^(A) is independently selected at each occurrence from:

-   -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³,        —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹,        —NRIC(O)N(R¹)₂, —C(O)N(R¹)₂, —C(O)NR²R³;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is        independently optionally substituted at each occurrence with one        or more substituents selected from —OR¹, —N(R¹)₂, —C(O)OR¹, C₃₋₈        carbocycle, and 3- to 10-membered heterocycle; and    -   C₃₋₈ carbocycle and 3- to 10-membered heterocycle,    -   wherein each C₃₋₈ carbocycle and 3- to 10-membered heterocycle        in R^(A) is independently optionally substituted with one or        more substituents selected from —OR¹, —CH₂N(R¹)₂, —N(R¹)₂,        —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —C(O)N(R¹)₂, R¹, and C₁₋₆ alkyl.

For a compound of Formula (I), (I-A), (I-B) or (I-C), R^(A) mayindependently be selected at each occurrence from:

-   -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂,        —C(O)N(R¹)₂;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is        independently optionally substituted at each occurrence with one        or more substituents selected from —OR¹, —N(R¹)₂, and 3- to        10-membered heterocycle; and    -   3- to 10-membered heterocycle,    -   wherein each 3- to 10-membered heterocycle in R^(A) is        independently optionally substituted with one or more        substituents selected from —OR¹, —N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹,        —C(O)OR¹, —C(O)N(R¹)₂, and R¹.

For a compound of Formula (I), (I-A), (I-B) or (I-C), R^(A) mayindependently be selected at each occurrence from:

-   -   —N(R¹)₂ and —NR¹C(O)R¹;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₆ alkyl, each of which is substituted        with 4- to 8-membered heterocycle; and    -   4- to 8-membered heterocycle,    -   wherein each 4- to 8-membered heterocycle in R^(A) is        independently optionally substituted with one or more        substituents selected from —C(O)OR¹ and R¹.

For a compound of Formula (I), (I-A), (I-B) or (I-C), R^(A) may be—N(R¹)₂. In some embodiments, R^(A) is —NR¹C(O)R¹. In some embodiments,R^(A) is C₁₋₆ alkyl substituted with 4- to 8-membered heterocycle,wherein the 4- to 8-membered heterocycle is substituted with —C(O)OR¹.In some embodiments, R^(A) is —N(R¹)—C₁_6 alkyl substituted with 4- to8-membered heterocycle. In some embodiments, R^(A) is 4- to 8-memberedheterocycle, wherein the 4- to 8-membered heterocycle is optionallysubstituted with one or more substituents selected from —C(O)OR¹ and R¹.

For a compound of Formula (I), (I-A), (I-B) or (I-C), R¹ mayindependently be selected at each occurrence from hydrogen; and C₁₋₆alkyl, 1- to 6-membered heteroalkyl, C₀₋₃ alkyl-(C_(3-s) carbocycle),and C₀₋₃ alkyl-(3- to 10-membered heterocycle), each of which isoptionally substituted by one or more substituents selected from —NH₂,—NHCH₃, —N(CH₃)₂, —CH₂CH₂N(CH₃)₂, —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH,—CH₂OH, —CH₂CH₂OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃,C₃₋₈ carbocycle, and 3- to 6-membered heterocycle. In some embodiments,R¹ is independently selected at each occurrence from hydrogen; and C₁₋₆alkyl and C₀₋₃ alkyl-(3- to 10-membered heterocycle), each of which isoptionally substituted by one or more substituents selected from —OH,—CH₂OH, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-memberedheterocycle.

For a compound of Formula (I), (I-A), (I-B) or (I-C), m may be 1 or 2,such as m is 1.

For a compound of Formula (I), (I-A), (I-B) or (I-C), B may be selectedfrom C₆₋₁₀ aryl and 5- to 10-membered heteroaryl. In some embodiments, Bis selected from phenyl and pyridyl. In some embodiments, B is pyridyl,such as 2-pyridyl.

For a compound of Formula (I), (I-A), (I-B) or (I-C), R^(B) mayindependently be selected at each occurrence from halogen, —NH₂, —NHCH₃,—NHCH₂CH₃, —C(O)CH₃, —OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂,—C(CH₃)₃, —CH₂F, —CHF₂, and —CF₃. In some embodiments, R^(B) isindependently selected at each occurrence from halogen, —OH, —CH₃, and—CHF₂, such as —CH₃.

For a compound of Formula (I), (I-A), (I-B) or (I-C), n may be 1 or 2,such as n is 1.

The compound of Formula (I) may be a compound of Formula (I-D):

The compound of Formula (I) may be a compound of Formula (I-E):

In some embodiments, for a compound of Formula (I):

-   -   A is selected from phenyl and 5- to 6-membered heteroaryl;    -   B is pyridyl;    -   R^(A) is independently selected at each occurrence from:        -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂,            —S(═O)₂NR²R³, —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹,            —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂,            —C(O)NR²R³;        -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is            independently optionally substituted at each occurrence with            one or more substituents selected from —OR¹, —N(R¹)₂,            —C(O)OR¹, C₃₋₈ carbocycle, and 3- to 10-membered            heterocycle; and        -   C₃₋₈ carbocycle and 3- to 10-membered heterocycle,        -   wherein each C₃₋₈ carbocycle and 3- to 10-membered            heterocycle in R^(A) is independently optionally substituted            with one or more substituents selected from —OR¹,            —CH₂N(R¹)₂, —N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹,            —C(O)N(R¹)₂, R¹, and C₁₋₆ alkyl;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl, 1- to 6-membered heteroalkyl, C₀₋₃        alkyl-(C_(3-s) carbocycle), and C₀₋₃ alkyl-(3- to 10-membered        heterocycle), each of which is optionally substituted by one or        more substituents selected from —NH₂, —NHCH₃, —N(CH₃)₂,        —CH₂CH₂N(CH₃)₂, —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH,        —CH₂CH₂OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃,        C₃₋₈ carbocycle, and 3- to 6-membered heterocycle;    -   m is 1 or 2;    -   R^(B) is selected from halogen, —OH, —CH₃, and —CHF₂; and    -   n is 1.

In some embodiments, for a compound of Formula (I):

-   -   A is phenyl;    -   B is pyridyl;    -   R^(A) is independently selected at each occurrence from:        -   —N(R¹)₂ and —NR¹C(O)R¹;        -   C₁₋₆ alkyl and —N(R¹)—C₁₋₆ alkyl, each of which is            substituted with 4- to 8-membered heterocycle; and        -   4- to 8-membered heterocycle,        -   wherein each 4- to 8-membered heterocycle in R^(A) is            independently optionally substituted with one or more            substituents selected from —C(O)OR¹ and R¹;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl and C₀₋₃ alkyl-(3- to 10-membered heterocycle),        each of which is optionally substituted by one or more        substituents selected from —OH, —CH₂OH, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-membered heterocycle;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In some embodiments, for a compound of Formula (I):

-   -   A is phenyl;    -   B is pyridyl;    -   R^(A) is C₁₋₆ alkyl substituted with 4- to 8-membered        heterocycle, wherein the 4- to 8-membered heterocycle is        substituted with —C(O)OR¹;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl and C₀₋₃ alkyl-(3- to 10-membered heterocycle),        each of which is optionally substituted by one or more        substituents selected from —OH, —CH₂OH, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-membered heterocycle;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In certain aspects, the present disclosure provides a substantially purestereoisomer of a compound disclosed herein. The stereoisomer may beprovided in at least 90% enantiomeric excess.

In certain aspects, the present disclosure provides a compound selectedfrom Table 1. In some embodiments, the present disclosure provides acompound selected from Table 2.

In certain aspects, the present disclosure provides a conjugate of theformula:

A′

L¹-D′)_(p),

wherein:

-   -   A′ is an antibody construct or targeting moiety;    -   L′ is a linker;    -   D′ is a compound or salt disclosed herein; and    -   p is an integer from 1 to 20

In certain aspects, the present disclosure provides a pharmaceuticalcomposition comprising a compound disclosed herein and apharmaceutically acceptable carrier. The pharmaceutical composition maybe formulated for inhalation.

In certain aspects, the present disclosure provides a method ofinhibiting ALK5, comprising contacting ALK5 with an effective amount ofa compound disclosed herein. In certain aspects, the present disclosureprovides a method of treating an ALK5-mediated disease or condition in asubject, comprising administering to the subject a therapeuticallyeffective amount of a compound disclosed herein. In practicing any ofthe subject methods, the disease or condition may be selected fromfibrosis, alopecia and cancer. In some embodiments, the disease orcondition is fibrosis. In some embodiments, the present disclosureprovides a method of treating fibrosis, comprising administering to apatient a therapeutically effective amount of a compound disclosedherein. The fibrosis may be selected from systemic sclerosis,nephrogenic systemic fibrosis, organ-specific fibrosis, fibrosisassociated with cancer, cystic fibrosis, and fibrosis associated with anautoimmune disease. Optionally, the organ-specific fibrosis is selectedfrom cardiac fibrosis, kidney fibrosis, pulmonary fibrosis, liverfibrosis, portal vein fibrosis, skin fibrosis, bladder fibrosis,intestinal fibrosis, peritoneal fibrosis, myelofibrosis, oral submucousfibrosis, and retinal fibrosis. In some embodiments, the organ-specificfibrosis is intestinal fibrosis. Optionally, the pulmonary fibrosis isselected from idiopathic pulmonary fibrosis (IPF), familial pulmonaryfibrosis (FPF), interstitial lung fibrosis, fibrosis associated withasthma, fibrosis associated with chronic obstructive pulmonary disease(COPD), silica-induced fibrosis, asbestos-induced fibrosis andchemotherapy-induced lung fibrosis. Optionally, the pulmonary fibrosisis idiopathic pulmonary fibrosis (IPF). In some embodiments, thepulmonary fibrosis was induced by a viral infection.

In practicing any of the subject methods, the disease or condition maybe cancer, optionally wherein the cancer is selected from breast cancer,colon cancer, prostate cancer, lung cancer, hepatocellular carcinoma,glioblastoma, melanoma, and pancreatic cancer. In some embodiments, thecancer is lung cancer, optionally non-small cell lung cancer. A methodof the subject disclosure may further comprise administering a secondtherapeutic agent. Optionally, the second therapeutic agent is animmunotherapeutic agent, such as a PD-1 inhibitor or a CTLA-4 inhibitor.In some embodiments, the immunotherapeutic agent is selected frompembrolizumab and durvalumab. A method of the present disclosure mayfurther comprise administering an effective amount of radiation. Inpracticing any of the subject methods, the compound or salt disclosedherein may be administered by inhalation.

In certain aspects, the present disclosure provides a compound disclosedherein for use in treating fibrosis. In certain aspects, the presentdisclosure provides the use of a compound disclosed herein for themanufacture of a medicament for treating fibrosis.

INCORPORATION BY REFERENCE

All publications and patent applications mentioned in this specificationare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

DETAILED DESCRIPTION

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which this disclosure belongs.

Chemical structures are named herein according to IUPAC conventions asimplemented in ChemDraw® software (Perkin Elmer, Inc., Cambridge, MA).

As used in the specification and claims, the singular forms “a”, “an”and “the” include plural references unless the context clearly dictatesotherwise.

The term “C_(x-y)” or “C_(x)-C_(y)” when used in conjunction with achemical moiety, such as alkyl, alkenyl, or alkynyl, is meant to includegroups that contain from x to y carbons in the chain. For example, theterm “C_(x-y) alkyl” refers to substituted or unsubstituted saturatedhydrocarbon groups, including straight-chain alkyl and branched-chainalkyl groups, that contain from x to y carbons in the chain.

“Alkyl” refers to substituted or unsubstituted saturated hydrocarbongroups, including linear and branched alkyl groups. An alkyl group maycontain from one to twelve carbon atoms (e.g., C₁₋₁₂ alkyl), such as oneto eight carbon atoms (C₁₋₈ alkyl) or one to six carbon atoms (C₁₋₆alkyl). Exemplary alkyl groups include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl,neopentyl, hexyl, septyl, octyl, nonyl, and decyl. An alkyl group isattached to the rest of the molecule by a single bond. Unless statedotherwise specifically in the specification, an alkyl group isoptionally substituted by one or more substituents such as thosesubstituents described herein.

“Haloalkyl” refers to an alkyl group that is substituted by one or morehalogens. Exemplary haloalkyl groups include trifluoromethyl,difluoromethyl, trichloro methyl, 2,2,2-trifluoroethyl,1,2-difluoroethyl, 3-bromo-2-fluoropropyl, and 1,2-dibromoethyl.

“Alkenyl” refers to substituted or unsubstituted hydrocarbon groups,including linear and branched alkenyl groups, containing at least onedouble bond. An alkenyl group may contain from two to twelve carbonatoms (e.g., C₂₋₁₂ alkenyl), such as two to eight carbon atoms (C₂₋₈alkenyl) or two to six carbon atoms (C₂₋₆ alkenyl). Exemplary alkenylgroups include ethenyl (i.e., vinyl), prop-1-enyl, but-1-enyl,pent-1-enyl, penta-1,4-dienyl, and the like. Unless stated otherwisespecifically in the specification, an alkenyl group is optionallysubstituted by one or more substituents such as those substituentsdescribed herein.

“Alkynyl” refers to substituted or unsubstituted hydrocarbon groups,including linear and branched alkynyl groups, containing at least onetriple bond. An alkynyl group may contain from two to twelve carbonatoms (e.g., C₂₋₁₂ alkynyl), such as two to eight carbon atoms (C₂₋₈alkynyl) or two to six carbon atoms (C₂₋₆ alkynyl). Exemplary alkynylgroups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and thelike. Unless stated otherwise specifically in the specification, analkynyl group is optionally substituted by one or more substituents suchas those substituents described herein.

“Alkylene” or “alkylene chain” refers to substituted or unsubstituteddivalent saturated hydrocarbon groups, including linear alkylene andbranched alkylene groups, that contain from one to twelve carbon atoms(e.g., C₁₋₁₂ alkylene), such as one to eight carbon atoms (C₁₋₈alkylene) or one to six carbon atoms (C₁₋₆ alkylene). Exemplary alkylenegroups include methylene, ethylene, propylene, and n-butylene.Similarly, “alkenylene” and “alkynylene” refer to alkylene groups, asdefined above, which comprise one or more carbon-carbon double or triplebonds, respectively. The points of attachment of the alkylene,alkenylene or alkynylene chain to the rest of the molecule can bethrough one carbon or any two carbons of the chain. Unless statedotherwise specifically in the specification, an alkylene, alkenylene, oralkynylene group is optionally substituted by one or more substituentssuch as those substituents described herein.

“Heteroalkyl”, “heteroalkenyl” and “heteroalkynyl” refer to substitutedor unsubstituted alkyl, alkenyl and alkynyl groups, respectively, inwhich one or more, such as 1, 2 or 3, of the carbon atoms are replacedwith a heteroatom, such as O, N, P, Si, S, or combinations thereof. Anynitrogen, phosphorus, and sulfur heteroatoms present in the chain mayoptionally be oxidized, and any nitrogen heteroatoms may optionally bequaternized. If given, a numerical range refers to the chain length intotal. For example, a 3- to 8-membered heteroalkyl group has a chainlength of 3 to 8 atoms. Connection to the rest of the molecule may bethrough either a heteroatom or a carbon in the heteroalkyl,heteroalkenyl or heteroalkynyl chain. Unless stated otherwisespecifically in the specification, a heteroalkyl, heteroalkenyl, orheteroalkynyl group is optionally substituted by one or moresubstituents such as those substituents described herein.

“Heteroalkylene”, “heteroalkenylene” and “heteroalkynylene” refer tosubstituted or unsubstituted alkylene, alkenylene and alkynylene groups,respectively, in which one or more, such as 1, 2 or 3, of the carbonatoms are replaced with a heteroatom, such as O, N, P, Si, S, orcombinations thereof. Any nitrogen, phosphorus, and sulfur heteroatomspresent in the chain may optionally be oxidized, and any nitrogenheteroatoms may optionally be quaternized. If given, a numerical rangerefers to the chain length in total. For example, a 3- to 8-memberedheteroalkylene group has a chain length of 3 to 8 atoms. The points ofattachment of the heteroalkylene, heteroalkenylene or heteroalkynylenechain to the rest of the molecule can be through either one heteroatomor one carbon, or any two heteroatoms, any two carbons, or any oneheteroatom and any one carbon in the heteroalkylene, heteroalkenylene orheteroalkynylene chain. Unless stated otherwise specifically in thespecification, a heteroalkylene, heteroalkenylene, or heteroalkynylenegroup is optionally substituted by one or more substituents such asthose substituents described herein.

“Carbocycle” refers to a saturated, unsaturated or aromatic ring inwhich each atom of the ring is a carbon atom. Carbocycle may includeC₃₋₁₀ monocyclic rings, C₆₋₁₂ bicyclic rings, C₆₋₁₂ spirocyclic rings,and C₆₋₁₂ bridged rings. Each ring of a bicyclic carbocycle may beselected from saturated, unsaturated, and aromatic rings. In someembodiments, the carbocycle is a C₆₋₁₂ aryl group, such as C₆₋₁₀ aryl.In some embodiments, the carbocycle is a C₆₋₁₂ cycloalkyl group. In someembodiments, the carbocycle is a C₆₋₁₂ cycloalkenyl group. In anexemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to asaturated or unsaturated ring, e.g., cyclohexane, cyclopentane, orcyclohexene. Any combination of saturated, unsaturated and aromaticbicyclic rings, as valence permits, are included in the definition ofcarbocycle. Exemplary carbocycles include cyclopentyl, cyclohexyl,cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. Unless statedotherwise specifically in the specification, a carbocycle is optionallysubstituted by one or more substituents such as those substituentsdescribed herein.

“Heterocycle” refers to a saturated, unsaturated or aromatic ringcomprising one or more heteroatoms, for example 1, 2 or 3 heteroatomsselected from O, S and N. Heterocycles include 3- to 10-memberedmonocyclic rings, 6- to 12-membered bicyclic rings, 6- to 12-memberedspirocyclic rings, and 6- to 12-membered bridged rings. Each ring of abicyclic heterocycle may be selected from saturated, unsaturated, andaromatic rings. The heterocycle may be attached to the rest of themolecule through any atom of the heterocycle, valence permitting, suchas a carbon or nitrogen atom of the heterocycle. In some embodiments,the heterocycle is a 5- to 10-membered heteroaryl group, such as 5- or6-membered heteroaryl. In some embodiments, the heterocycle is a 3- to12-membered heterocycloalkyl group. In an exemplary embodiment, aheterocycle, e.g., pyridyl, may be fused to a saturated or unsaturatedring, e.g., cyclohexane, cyclopentane, or cyclohexene. Exemplaryheterocycles include pyrrolidinyl, pyrrolyl, imidazolyl, pyrazolyl,triazolyl, piperidinyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl,thiophenyl, oxazolyl, thiazolyl, morpholinyl, indazolyl, indolyl, andquinolinyl. Unless stated otherwise specifically in the specification, aheterocycle is optionally substituted by one or more substituents suchas those substituents described herein.

“Heteroaryl” refers to a 5- to 12-membered aromatic ring that comprisesat least one heteroatom, such as 1, 2 or 3 heteroatoms, selected from O,S and N. As used herein, the heteroaryl ring may be selected frommonocyclic or bicyclic-including fused, spirocyclic and bridged ringsystems—wherein at least one of the rings in the ring system isaromatic. The heteroatom(s) in the heteroaryl may optionally beoxidized. One or more nitrogen atoms, if present, are optionallyquaternized. The heteroaryl may be attached to the rest of the moleculethrough any atom of the heteroaryl, valence permitting, such as a carbonor nitrogen atom of the heteroaryl. Examples of heteroaryl groupsinclude, but are not limited to, azepinyl, benzimidazolyl,benzisothiazolyl, benzisoxazolyl, benzofuranyl, benzothiazolyl,benzothiophenyl, benzoxazolyl, furanyl, imidazolyl, indazolyl, indolyl,isoquinolinyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purinyl,pyrazinyl, pyrazolidinyl, pyrazolyl, pyridazinyl, pyridazolyl, pyridyl,pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl,tetrahydroquinolinyl, thiadiazolyl, thiazolyl, and thienyl groups.Unless stated otherwise specifically in the specification, a heteroarylis optionally substituted by one or more substituents such as thosesubstituents described herein.

Unless stated otherwise, hydrogen atoms are implied in structuresdepicted herein as necessary to satisfy the valence requirement.

A waved line “

” drawn across a bond or a dashed bond ‘

’ are used interchangeably herein to denote where a bond disconnectionor attachment occurs. For example, in the structure

R^(a) is attached to the para position of a fluorophenyl ring through asingle bond. If R^(a) is 2-pyridine as in

then R^(a) may be depicted as

The term “substituted” refers to moieties having substituents replacinga hydrogen on one or more carbons or heteroatoms of the structure. Itwill be understood that “substitution” or “substituted with” includesthe implicit proviso that such substitution is in accordance withpermitted valence of the substituted atom and the substituent, and thatthe substitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc. As used herein, the term “substituted” iscontemplated to include all permissible substituents of organiccompounds. In a broad aspect, the permissible substituents includeacyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and non-aromatic substituents of organiccompounds. The permissible substituents can be one or more and the sameor different for appropriate organic compounds. For purposes of thisdisclosure, heteroatoms such as nitrogen may have any permissiblesubstituents of organic compounds described herein which satisfy thevalences of the heteroatoms.

A compound disclosed herein, such as a compound of Formula (I), isoptionally substituted by one or more, such as 1, 2 or 3 substituentsselected from:

-   -   halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³, —S(═O)R¹,        —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —C(O)OR¹, —OC(O)R¹,        —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹, —NR¹C(O)OR¹,        —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂, —C(O)NR²R³,        —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S, ═N(R¹);    -   C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, and —N(R¹)—C₁₋₁₀        alkyl, each of which is independently optionally substituted at        each occurrence with one or more substituents selected from        halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³, —S(═O)R¹,        —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —C(O)OR¹, —OC(O)R¹,        —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹, —NR¹C(O)OR¹,        —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂, —C(O)NR²R³,        —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S, ═N(R¹), C₃₋₁₂ carbocycle, and        3- to 12-membered heterocycle; and    -   C₃₋₁₂ carbocycle and 3- to 12-membered heterocycle,    -   wherein each C₃₋₁₂ carbocycle and 3- to 12-membered heterocycle        is independently optionally substituted with one or more        substituents selected from halogen, —NO₂, —CN, —OR¹, —SR¹,        —CH₂N(R¹)₂, —N(R¹)₂, —NR²R³, —S(═O)R¹, —S(═O)₂R¹, —S(═O)₂N(R¹)₂,        —S(═O)₂NR²R³, —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³,        —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —OC(O)R¹, —OC(O)OR¹,        —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹, —NR¹C(O)OR¹,        —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂, —C(O)NR²R³,        —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S, ═N(R¹), R¹, C₁₋₆ alkyl, C₁₋₆        haloalkyl, C₂₋₆ alkenyl, and C₂₋₆ alkynyl;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, 1- to 6-membered        heteroalkyl, C₀₋₃ alkyl-(C₃₋₁₂ carbocycle), and C₀₋₃ alkyl-(3-        to 12-membered heterocycle), each of which is optionally        substituted by one or more substituents selected from halogen,        —CN, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —CH₂CH₂N(CH₃)₂,        —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃,        —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₁₂ carbocycle,        and 3- to 6-membered heterocycle; and    -   R² and R³ are taken together with the nitrogen atom to which        they are attached to form a heterocycle, optionally substituted        with one or more R¹.

In some embodiments, a compound disclosed herein, such as a compound ofFormula (I), is optionally substituted by one or more, such as 1, 2 or 3substituents selected from: halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂,—NR²R³, —S(═O)R¹, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,—NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —C(O)OR¹, —OC(O)R¹,—OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹, —NR¹C(O)OR¹,—NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂, —C(O)NR²R³, —P(O)(OR¹)₂,—P(O)(R¹)₂, ═O, ═S, ═N(R¹);

-   -   C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, and —N(R¹)—C₁₋₁₀        alkyl, each of which is independently optionally substituted at        each occurrence with one or more substituents selected from        halogen, ═O, ═S, ═N(R¹), C₃₋₁₂ carbocycle, and 3- to 12-membered        heterocycle; and    -   C₃₋₁₂ carbocycle and 3- to 12-membered heterocycle,    -   wherein each C₃₋₁₂ carbocycle and 3- to 12-membered heterocycle        is independently optionally substituted with one or more        substituents selected from halogen, ═O, ═S, ═N(R¹), C₁₋₆ alkyl,        C₁₋₆ haloalkyl, C₂₋₆ alkenyl, and C₂₋₆ alkynyl;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, 1- to 6-membered        heteroalkyl, C₀₋₃ alkyl-(C₃₋₁₂ carbocycle), and C₀₋₃ alkyl-(3-        to 12-membered heterocycle), each of which is optionally        substituted by one or more substituents selected from halogen,        —CN, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —CH₂CH₂N(CH₃)₂,        —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃,        —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₁₂ carbocycle,        and 3- to 6-membered heterocycle; and    -   R² and R³ are taken together with the nitrogen atom to which        they are attached to form a heterocycle, optionally substituted        with one or more R¹.

In some embodiments, a compound disclosed herein, such as a compound ofFormula (I), is optionally substituted by one or more, such as 1, 2 or 3substituents selected from halogen, —CN, —NO₂, —NH₂, —NHCH₃, —NHCH₂CH₃,═O, —OH, —OCH₃, and —OCH₂CH₃; and C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, 1- to 6-membered heteroalkyl, C₃₋₁₂ carbocycle, and 3- to12-membered heterocycle, each of which is optionally substituted byhalogen, —CN, —NO₂, —NH₂, —NHCH₃, —NHCH₂CH₃, ═O, —OH, —OCH₃, —OCH₂CH₃,—CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₁₂ carbocycle, or 3- to6-membered heterocycle.

It will be understood by those skilled in the art that substituents canthemselves be substituted, if appropriate. Unless specifically stated as“unsubstituted”, references to chemical moieties herein are understoodto include substituted variants. For example, reference to a“heteroaryl” group or moiety implicitly includes both substituted andunsubstituted variants.

Where bivalent substituent groups are specified herein by theirconventional chemical formulae, written from left to right, they areintended to encompass the isomer that would result from writing thestructure from right to left, e.g., —CH₂O— is also intended to encompassto —OCH₂—.

“Optional” or “optionally” means that the subsequently described eventof circumstances may or may not occur, and that the description includesinstances where the event or circumstance occurs and instances in whichit does not. For example, an “optionally substituted” group may beeither unsubstituted or substituted.

Compounds of the present disclosure also include crystalline andamorphous forms of those compounds, pharmaceutically acceptable salts,and active metabolites having the same type of activity, including, forexample, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvatedpolymorphs (including anhydrates), conformational polymorphs, amorphousforms of the compounds, and mixtures thereof.

The compounds described herein may exhibit their natural isotopicabundance, or one or more of the atoms may be artificially enriched in aparticular isotope having the same atomic number, but an atomic mass ormass number different from the atomic mass or mass number predominantlyfound in nature. All isotopic variations of the compounds of the presentdisclosure, whether radioactive or not, are encompassed within the scopeof the present disclosure. For example, hydrogen has three naturallyoccurring isotopes, denoted ¹H (protium), ²H (deuterium), and ³H(tritium). Protium is the most abundant isotope of hydrogen in nature.Enriching for deuterium may afford certain therapeutic advantages, suchas increased in vivo half-life and/or exposure, or may provide acompound useful for investigating in vivo routes of drug elimination andmetabolism. Examples of isotopes that may be incorporated into compoundsof the present disclosure include, but are not limited to, ²H, ³H, ¹³C,¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ³⁶Cl, and ¹⁸F. Of particular interest arecompounds of Formula (I) enriched in tritium or carbon-14, which can beused, for example, in tissue distribution studies; compounds of thedisclosure enriched in deuterium especially at a site of metabolism,resulting, for example, in compounds having greater metabolic stability;and compounds of Formula (I) enriched in a positron emitting isotope,such as ¹¹C, ¹⁸F, ¹⁵O and ¹³N, which can be used, for example, inPositron Emission Topography (PET) studies. Isotopically-enrichedcompounds may be prepared by conventional techniques well known to thoseskilled in the art.

As used herein, the phrase “of the formula” or “having the formula” or“having the structure” is not intended to be limiting and is used in thesame way that the term “comprising” is commonly used. For example, ifone structure is depicted, it is understood that all stereoisomer andtautomer forms are encompassed, unless stated otherwise.

Certain compounds described herein contain one or more asymmetriccenters and can thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms, the asymmetric centers of which can be defined, interms of absolute stereochemistry, as (R)- or (S)-. In some embodiments,in order to optimize the therapeutic activity of the compounds of thedisclosure, e.g., to treat fibrosis, it may be desirable that the carbonatoms have a particular configuration (e.g., (R,R), (S,S), (S,R), or(R,S)) or are enriched in a stereoisomeric form having suchconfiguration. The compounds of the disclosure may be provided asracemic mixtures. Accordingly, the disclosure relates to racemicmixtures, pure stereoisomers (e.g., enantiomers and diastereoisomers),stereoisomer-enriched mixtures, and the like, unless otherwiseindicated. When a chemical structure is depicted herein without anystereochemistry, it is understood that all possible stereoisomers areencompassed by such structure. Similarly, when a particular stereoisomeris shown or named herein, it will be understood by those skilled in theart that minor amounts of other stereoisomers may be present in thecompositions of the disclosure unless otherwise indicated, provided thatthe utility of the composition as a whole is not eliminated by thepresence of such other isomers. Individual stereoisomers may be obtainedby numerous methods that are known in the art, including preparationusing chiral synthons or chiral reagents, resolution using chiralchromatography using a suitable chiral stationary phase or support, orby chemically converting them into diastereoisomers, separating thediastereoisomers by conventional means such as chromatography orrecrystallization, then regenerating the original stereoisomer.

Additionally, where applicable, all cis-trans or E/Z isomers (geometricisomers), tautomeric forms and topoisomeric forms of the compounds ofthe invention are included within the scope of the invention unlessotherwise specified.

The term “tautomer”, as used herein, refers to each of two or moreisomers of a compound that exist in equilibrium and which readyinterconvert. For example, one skilled in the art would readilyunderstand that 1,2,3-triazole exists in two tautomeric forms:

Unless otherwise specified, chemical entities described herein areintended to include all possible tautomers, even when a structuredepicts only one of them.

The term “pharmaceutically acceptable” refers to a material that is notbiologically or otherwise unacceptable when used in the subjectcompositions and methods. For example, the term “pharmaceuticallyacceptable carrier” refers to a material—such as an adjuvant, excipient,glidant, sweetening agent, diluent, preservative, dye, colorant, flavorenhancer, surfactant, wetting agent, dispersing agent, suspending agent,stabilizer, isotonic agent, solvent or emulsifier—that can beincorporated into a composition and administered to a patient withoutcausing unacceptable biological effects or interacting in anunacceptable manner with other components of the composition. Suchpharmaceutically acceptable materials typically have met the requiredstandards of toxicological and manufacturing testing, and include thosematerials identified as suitable inactive ingredients by the U.S. Foodand Drug Administration.

The terms “salt” and “pharmaceutically acceptable salt” refer to a saltprepared from a base or an acid. Pharmaceutically acceptable salts aresuitable for administration to a patient, such as a mammal (for example,salts having acceptable mammalian safety for a given dosage regime).Salts can be formed from inorganic bases, organic bases, inorganic acidsand organic acids. In addition, when a compound contains both a basicmoiety, such as an amine, pyridine or imidazole, and an acidic moiety,such as a carboxylic acid or tetrazole, zwitterions may be formed andare included within the term “salt” as used herein. Salts derived frominorganic bases include ammonium, calcium, copper, ferric, ferrous,lithium, magnesium, manganic, manganous, potassium, sodium, and zincsalts, and the like. Salts derived from organic bases include salts ofprimary, secondary and tertiary amines, including substituted amines,cyclic amines, naturally-occurring amines and the like, such asarginine, betaine, caffeine, choline, N,N′-dibenzylethylenediamine,diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol,ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine,glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. Salts derived from inorganicacids include salts of boric, carbonic, hydrohalic (hydrobromic,hydrochloric, hydrofluoric or hydroiodic), nitric, phosphoric, sulfamicand sulfuric acids. Salts derived from organic acids include salts ofaliphatic hydroxyl acids (for example, citric, gluconic, glycolic,lactic, lactobionic, malic, and tartaric acids), aliphaticmonocarboxylic acids (for example, acetic, butyric, formic, propionicand trifluoroacetic acids), amino acids (for example, aspartic andglutamic acids), aromatic carboxylic acids (for example, benzoic,p-chlorobenzoic, diphenylacetic, gentisic, hippuric, and triphenylaceticacids), aromatic hydroxyl acids (for example, o-hydroxybenzoic,p-hydroxybenzoic, 1-hydroxynaphthalene-2-carboxylic and3-hydroxynaphthalene-2-carboxylic acids), ascorbic, dicarboxylic acids(for example, fumaric, maleic, oxalic and succinic acids), glucoronic,mandelic, mucic, nicotinic, orotic, pamoic, pantothenic, sulfonic acids(for example, benzenesulfonic, camphorsulfonic, edisylic,ethanesulfonic, isethionic, methanesulfonic, naphthalenesulfonic,naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic andp-toluenesulfonic acids), xinafoic acid, and the like.

The term “therapeutically effective amount” refers to that amount of acompound described herein that is sufficient to affect treatment whenadministered to a subject in need thereof. For example, atherapeutically effective amount for treating pulmonary fibrosis is anamount of compound needed to, for example, reduce, suppress, eliminate,or prevent the formation of fibrosis in a subject, or to treat theunderlying cause of pulmonary fibrosis. The therapeutically effectiveamount may vary depending upon the intended treatment application (invivo), or the subject and disease condition being treated, e.g., theweight and age of the subject, the severity of the disease condition,the manner of administration and the like, which can readily bedetermined by one of ordinary skill in the art. The specific dose willvary depending on the particular compound chosen, the dosing regimen tobe followed, whether it is administered in combination with othercompounds, timing of administration, the tissue to which it isadministered, and the physical delivery system in which it is carried.The term “effective amount” refers to an amount sufficient to obtain adesired result, which may not necessarily be a therapeutic result. Forexample, an “effective amount” may be the amount needed to inhibit anenzyme.

As used herein, “treating” or “treatment” refers to an approach forobtaining beneficial or desired results with respect to a disease,disorder, or medical condition (such as pulmonary fibrosis) in asubject, including but not limited to the following: (a) preventing thedisease or medical condition from occurring, e.g., preventing thereoccurrence of the disease or medical condition or prophylactictreatment of a subject that is pre-disposed to the disease or medicalcondition; (b) ameliorating the disease or medical condition, e.g.,eliminating or causing regression of the disease or medical condition ina subject; (c) suppressing the disease or medical condition, e.g.,slowing or arresting the development of the disease or medical conditionin a subject; or (d) alleviating symptoms of the disease or medicalcondition in a subject. For example, “treating pulmonary fibrosis” wouldinclude preventing fibrosis from occurring, ameliorating fibrosis,suppressing fibrosis, and alleviating the symptoms of fibrosis (forexample, increasing oxygen levels in blood or improved lung functiontests). Also, a therapeutic benefit is achieved with the eradication oramelioration of one or more of the physiological symptoms associatedwith the underlying disorder such that an improvement is observed in thesubject, notwithstanding that the subject may still be afflicted withthe underlying disorder

A “therapeutic effect”, as that term is used herein, encompasses atherapeutic benefit and/or a prophylactic benefit as described above. Aprophylactic effect includes delaying or eliminating the appearance of adisease or condition, delaying or eliminating the onset of symptoms of adisease or condition, slowing, halting, or reversing the progression ofa disease or condition, or any combination thereof.

The terms “antagonist” and “inhibitor” are used interchangeably, andthey refer to a compound having the ability to inhibit a biologicalfunction (e.g., activity, expression, binding, protein-proteininteraction) of a target protein (e.g., ALK5). Accordingly, the terms“antagonist” and “inhibitor” are defined in the context of thebiological role of the target protein. While preferred antagonistsherein specifically interact with (e.g., bind to) the target, compoundsthat inhibit a biological activity of the target protein by interactingwith other members of the signal transduction pathway of which thetarget protein is a member are also specifically included within thisdefinition.

The term “selective inhibition” or “selectively inhibit” refers to theability of a biologically active agent to preferentially reduce thetarget signaling activity as compared to off-target signaling activity,via direct or indirect interaction with the target.

As used herein, the term “antibody” refers to an immunoglobulin moleculethat specifically binds to, or is immunologically reactive toward, aspecific antigen. An antibody may be, for example, polyclonal,monoclonal, genetically engineered, or an antigen binding fragmentthereof, and further may be, for example, murine, chimeric, humanized,heteroconjugate, bispecific, a diabody, a triabody, or a tetrabody. Anantigen binding fragment includes an antigen binding domain and may bein the form of, for example, a Fab′, F(ab′)₂, Fab, Fv, rIgG, scFv, hcAbs(heavy chain antibodies), a single domain antibody, V_(HH), V_(NAR),sdAb, or nanobody.

The term “antigen binding domain” as used herein refers to a region of amolecule that binds to an antigen. An antigen binding domain may be anantigen-binding portion of an antibody or an antibody fragment. Anantigen binding domain may be one or more fragments of an antibody thatretain the ability to specifically bind to an antigen. An antigenbinding domain can be an antigen binding fragment and may recognize asingle antigen, two antigens, three antigens or more. As used herein,“recognize” with regard to antibody interactions refers to theassociation or binding between an antigen binding domain of an antibodyor portion thereof and an antigen.

An “antibody construct” refers to a molecule, e.g., a protein, peptide,antibody or portion thereof, that contains an antigen binding domain andan Fc domain (e.g., an Fc domain from within the Fc region). An antibodyconstruct may recognize, for example, one antigen or multiple antigens.

A “targeting moiety” refers to a structure that has a selective affinityfor a target molecule relative to other non-target molecules. Thetargeting moiety binds to a target molecule. A targeting moiety mayinclude an antibody, a peptide, a ligand, a receptor, or a bindingportion thereof. The target biological molecule may be a biologicalreceptor or other structure of a cell, such as a tumor antigen.

The terms “subject” and “patient” refer to an animal, such as a mammal,for example a human. The methods described herein can be useful in bothhuman therapeutics and veterinary applications. In some embodiments, thesubject is a mammal, and in some embodiments, the subject is human.“Mammal” includes humans and both domestic animals such as laboratoryanimals and household pets (e.g., cats, dogs, swine, cattle, sheep,goats, horses, rabbits), and non-domestic animals such as wildlife andthe like.

“Prodrug” is meant to indicate a compound that may be converted underphysiological conditions or by solvolysis to a biologically activecompound described herein (e.g., compound of Formula (I), (I′), (I-A),(I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E)).Thus, the term “prodrug” refers to a precursor of a biologically activecompound that is pharmaceutically acceptable. In some aspects, a prodrugis inactive when administered to a subject but is converted in vivo toan active compound, for example, by hydrolysis. The prodrug compoundoften offers advantages of solubility, tissue compatibility or delayedrelease in a mammalian organism (see, e.g., Bundgard, H., Design ofProdrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam); Higuchi, T., etal., “Pro-drugs as Novel Delivery Systems,” (1987) A.C.S. SymposiumSeries, Vol. 14; and Bioreversible Carriers in Drug Design, ed. EdwardB. Roche, American Pharmaceutical Association and Pergamon Press) eachof which is incorporated in full by reference herein. The term “prodrug”is also meant to include any covalently bonded carriers, which releasethe active compound in vivo when such prodrug is administered to amammalian subject. Prodrugs of an active compound, as described herein,are typically prepared by modifying functional groups present in theactive compound in such a way that the modifications are cleaved, eitherin routine manipulation or in vivo, to the parent active compound.Prodrugs include compounds wherein a hydroxy, amino or mercapto group isbonded to any group that, when the prodrug of the active compound isadministered to a mammalian subject, cleaves to form a free hydroxy,free amino or free mercapto group, respectively. Examples of prodrugsinclude, but are not limited to, acetate, formate and benzoatederivatives of a hydroxy functional group, or acetamide, formamide andbenzamide derivatives of an amine functional group in the activecompound, and the like.

The term “in vivo” refers to an event that takes place in a subject'sbody.

The term “in vitro” refers to an event that takes places outside of asubject's body. For example, an in vitro assay encompasses any assay runoutside of a subject. In vitro assays encompass cell-based assays inwhich cells alive or dead are employed. In vitro assays also encompass acell-free assay in which no intact cells are employed.

The disclosure is also meant to encompass the in vivo metabolic productsof the disclosed compounds. Such products may result from, for example,the oxidation, reduction, hydrolysis, amidation, esterification, and thelike of the administered compound, primarily due to enzymatic processes.Accordingly, the disclosure includes compounds produced by a processcomprising administering a compound of this disclosure to a mammal for aperiod of time sufficient to yield a metabolic product thereof. Suchproducts are typically identified by administering a radiolabeledcompound of the disclosure in a detectable dose to an animal, such asrat, mouse, guinea pig, monkey, or to a human, allowing sufficient timefor metabolism to occur, and isolating its conversion products from theurine, blood or other biological samples.

Lung function tests include tests to check how well the lungs work.Spirometry, for example, measures the amount of air the lungs can holdas well as how forcefully one can empty air from the lungs. Forcedexpiratory volume (FEV) is a measure of the amount of air a person canexhale during a forced breath. FEV1, for example, is the amount of air aperson can force from their lungs in one second. Forced vital capacity(FVC) is the total amount of air exhaled during an FEV test. The ratioof FEV1/FVC, also known as Index of Air Flow or Tiffeneau-Pinelli Index,is a measurement used to assess the health of a patient's lung function.A ratio of <80% indicates an obstructive defect is present in the lungs,such as chronic obstructive pulmonary disease (COPD). A ratio of >80%indicates a restrictive defect is present in the lungs, such aspulmonary fibrosis. The ratio of >80% in restrictive lung diseaseresults from both FEV1 and FVC being reduced but that the decline in FVCis more than that of FEV1, resulting in a higher than 80% value.

The term “transforming growth factor-β” may also be referred to asTGF-β, transforming growth factor beta-1, or TGF-beta-1. It is alsocleaved into latency-associated peptide (LAP).

The term “TGF-β receptor II” may also be referred to as TβRII, type IITGF-β receptor, TGF-βRII, TGF-beta receptor type-2, TGFR-2, TGF-betatype II receptor, transforming growth factor-beta receptor type II,TGF-beta receptor type II or TbetaR-II.

The term “TGF-β receptor I” may also be referred to as TβRI, type ITGF-β receptor, TGF-βRI, TGF-beta receptor type-1, TGFR-1, activin Areceptor type II-like protein kinase of 53 kD, activin receptor-likekinase 5, ALK-5, ALK5, serine/threonine-protein kinase receptor R4,SKR4, TGF-beta type I receptor, transforming growth factor-beta receptortype I, TGF-beta receptor type I, transforming growth factor betareceptor I, TGF-beta receptor 1, or TbetaR-I.

The present disclosure provides compounds that are capable ofselectively binding to and/or modulating ALK5. In some embodiments, thecompounds modulate ALK5 by binding to or interacting with one or moreamino acids and/or one or more metal ions. The binding of thesecompounds may disrupt ALK5 downstream signaling.

In certain aspects, the present disclosure provides a compound ofFormula (I′):

or a pharmaceutically acceptable salt thereof, wherein:

-   -   A and B are each independently selected from C₃₋₁₂ carbocycle        and 3- to 12-membered heterocycle;    -   X¹ is selected from CH, CR^(D), N and S;    -   X² and X³ are each independently selected from CH, CR^(E), N and        S;    -   X⁴ is selected from bond, CH, CR^(E), N and S, provided that no        more than one of X², X³ and X⁴ is S;    -   R^(A), R^(B), R^(C), R^(D) and R^(E) are each independently        selected at each occurrence from:        -   halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³, —S(═O)R¹,            —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,            —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —C(O)OR¹,            —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹,            —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂,            —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S, ═N(R¹);        -   C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, and —N(R¹)—C₁₋₁₀            alkyl, each of which is independently optionally substituted            at each occurrence with one or more substituents selected            from halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³,            —S(═O)R¹, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³,            —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹,            —C(O)OR¹, —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³,            —NR¹C(O)R¹, —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³,            —C(O)N(R¹)₂, —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S,            ═N(R¹), C₃₋₁₂ carbocycle, and 3- to 12-membered heterocycle;            and        -   C₃₋₁₂ carbocycle and 3- to 12-membered heterocycle,        -   wherein each C₃₋₁₂ carbocycle and 3- to 12-membered            heterocycle in R^(A), R^(B), R^(C), R^(D) and R^(E) is            independently optionally substituted with one or more            substituents selected from halogen, —NO₂, —CN, —OR¹, —SR¹,            —CH₂N(R¹)₂, —N(R¹)₂, —NR²R³, —S(═O)R¹, —S(═O)₂R¹,            —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂,            —NR¹S(═O)₂NR²R³, —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —OC(O)R¹,            —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹,            —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂,            —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S, ═N(R¹), R¹,            C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, and C₂₋₆ alkynyl;        -   or two R^(B) groups, two R^(C) groups or two R^(E) groups            can together optionally form a bridge or ring;    -   m and n are each independently an integer from 0 to 3;    -   p is an integer from 0 to 2;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, 1- to 6-membered        heteroalkyl, C₀₋₃ alkyl-(C₃ 1₂ carbocycle), and C₀₋₃ alkyl-(3-        to 12-membered heterocycle), each of which is optionally        substituted by one or more substituents selected from halogen,        —CN, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —CH₂CH₂N(CH₃)₂,        —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃,        —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₁₂ carbocycle,        and 3- to 6-membered heterocycle; and    -   R² and R³ are taken together with the nitrogen atom to which        they are attached to form a heterocycle, optionally substituted        with one or more R¹.

In some embodiments, for a compound of Formula (I′), R^(B), R^(C), R^(D)and R^(E) are each independently selected at each occurrence fromhalogen, —CN, —NH₂, —NHCH₃, —NHCH₂CH₃, —C(O)CH₃, —OH, —OCH₃, —OCH₂CH₃,—CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂F, —CHF₂, —CF₃, C₃₋₄ carbocycle,and 3- to 4-membered heterocycle; or two R^(C) groups or two R^(E)groups can together optionally form a bridge or ring;

In some embodiments, for a compound of Formula (I′), p is 0 or 1, suchas p is 0. In some embodiments, R^(C) is selected from halogen, —NH₂ and—CH₃. In some embodiments, X¹ is selected from CH and N, such as X¹ isN. In some embodiments, X² is selected from CH, CR^(E), N and S, such asX² is N. In some embodiments, X² is selected from CH and CR^(E), whereinR^(E) is selected from —CN, —OCH₃ and —CH₃. In some embodiments, X³ isselected from CH and CR^(E), such as X³ is CH. In some embodiments, X³is CR^(E), wherein R^(E) is —OCH₃.

In some embodiments, X⁴ is selected from bond, CH, CR^(E) and N, such asX⁴ is CH. In some embodiments, X⁴ is N. In some embodiments, X³ and X⁴are each independently CR^(E), wherein the two R^(E) groups form a ring,such as a 5- or 6-membered aryl or heteroaryl ring. Non-limitingexamples wherein X³ and X⁴ are each independently CR^(E), wherein thetwo R^(E) groups form a ring include:

In some embodiments, X⁴ is a bond

Non-limiting examples wherein X⁴ is a bond include:

In some embodiments, the compound of Formula (I′) is a compound ofFormula (I′-A), (I′-B), (I′-C), (I′-D) or (I′-E):

In certain aspects, the present disclosure provides a compound ofFormula (I):

or a pharmaceutically acceptable salt thereof, wherein:

-   -   A and B are each independently selected from C₃₋₁₂ carbocycle        and 3- to 12-membered heterocycle;    -   R^(A) is independently selected at each occurrence from:        -   halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³, —S(═O)R¹,            —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,            —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —C(O)OR¹,            —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³, —NR¹C(O)R¹,            —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³, —C(O)N(R¹)₂,            —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S, ═N(R¹);        -   C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, and —N(R¹)—C₁₋₁₀            alkyl, each of which is independently optionally substituted            at each occurrence with one or more substituents selected            from halogen, —NO₂, —CN, —OR¹, —SR¹, —N(R¹)₂, —NR²R³,            —S(═O)R¹, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³,            —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹,            —C(O)OR¹, —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R¹)₂, —OC(O)NR²R³,            —NR¹C(O)R¹, —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³,            —C(O)N(R¹)₂, —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S,            ═N(R¹), C₃₋₁₂ carbocycle, and 3- to 12-membered heterocycle;            and        -   C₃₋₁₂ carbocycle and 3- to 12-membered heterocycle,        -   wherein each C₃₋₁₂ carbocycle and 3- to 12-membered            heterocycle in R^(A) is independently optionally substituted            with one or more substituents selected from halogen, —NO₂,            —CN, —OR¹, —SR¹, —CH₂N(R¹)₂, —N(R¹)₂, —NR²R³, —S(═O)R¹,            —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,            —NR¹S(═O)₂N(R¹)₂, —NR¹S(═O)₂NR²R³, —C(O)R¹, —CH₂C(O)OR¹,            —C(O)OR¹, —OC(O)R¹, —OC(O)OR¹, —OC(O)N(R)₂, —OC(O)NR²R³,            —NR¹C(O)R¹, —NR¹C(O)OR¹, —NR¹C(O)N(R¹)₂, —NR¹C(O)NR²R³,            —C(O)N(R¹)₂, —C(O)NR²R³, —P(O)(OR¹)₂, —P(O)(R¹)₂, ═O, ═S,            ═N(R¹), R¹, C₁_6 alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, and            C₂₋₆alkynyl;    -   R^(B) is independently selected at each occurrence from halogen,        —CN, —NH₂, —NHCH₃, —NHCH₂CH₃, —C(O)CH₃, —OH, —OCH₃, —OCH₂CH₃,        —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂F, —CHF₂, —CF₃, C₃₋₄        carbocycle, and 3- to 4-membered heterocycle;    -   m and n are each independently an integer from 0 to 3;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, 1- to 6-membered        heteroalkyl, C₀₋₃ alkyl-(C₃₋₁₂ carbocycle), and C₀₋₃ alkyl-(3-        to 12-membered heterocycle), each of which is optionally        substituted by one or more substituents selected from halogen,        —CN, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —CH₂CH₂N(CH₃)₂,        —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃,        —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₁₂ carbocycle,        and 3- to 6-membered heterocycle; and    -   R² and R³ are taken together with the nitrogen atom to which        they are attached to form a heterocycle, optionally substituted        with one or more R¹.

In some embodiments, the compound of Formula (I) is a compound ofFormula (I-A), (I-B), (I-C), (I-D) or (I-E):

In some embodiments, for a compound of Formula (I), (I′), (I-A) or(I′-A), A is selected from C₆₋₁₀ aryl and 5- to 10-membered heteroaryl.In some embodiments, A is selected from phenyl and 5- to 6-memberedheteroaryl, such as A is phenyl. In some embodiments, A is selected fromphenyl, indanyl, thiazolyl, thiophenyl, pyrazolyl, pyridyl, pyrimidinyl,indazolyl, benzotriazolyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, benzodioxanyl, and tetrahydrobenzazepinyl. Insome embodiments, A is selected from phenyl, pyridinyl, thiazolyl andthiophenyl. In some embodiments, A is phenyl. In some embodiments, A ispyridinyl. In some embodiments, A is thiazolyl. In some embodiments, Ais thiophenyl.

In some embodiments, the compound of Formula (I) is a compound of theformula:

In some embodiments, for a compound of Formula (I), (I′), (I-A) or(I′-A), m is 1 or 2. In some embodiments, m is 1. In some embodiments, Ais C₆ aryl or 6-membered heteroaryl, m is 1 and R^(A) is in the meta- orpara-position.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E), R^(A) isindependently selected at each occurrence from:

-   -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³,        —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹,        —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂, —C(O)NR²R³;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is        independently optionally substituted at each occurrence with one        or more substituents selected from halogen, —OR¹, —N(R¹)₂,        —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂,        —C(O)N(R¹)₂, —C(O)NR²R³, C₃₋₈ carbocycle, and 3- to 10-membered        heterocycle; and    -   C₃₋₈ carbocycle and 3- to 10-membered heterocycle,    -   wherein each C₃₋₈ carbocycle and 3- to 10-membered heterocycle        in R^(A) is independently optionally substituted with one or        more substituents selected from halogen, —OR¹, —CH₂N(R¹)₂,        —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —NR¹C(O)R¹,        —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂, —C(O)NR²R³, R¹, and C₁₋₆ alkyl.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E), R^(A) isindependently selected at each occurrence from:

-   -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³,        —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹,        —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂, —C(O)NR²R³;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is        independently optionally substituted at each occurrence with one        or more substituents selected from —OR¹, —N(R¹)₂, —C(O)OR¹, C₃₋₈        carbocycle, and 3- to 10-membered heterocycle; and    -   C₃₋₈ carbocycle and 3- to 10-membered heterocycle,    -   wherein each C₃₋₈ carbocycle and 3- to 10-membered heterocycle        in R^(A) is independently optionally substituted with one or        more substituents selected from —OR¹, —CH₂N(R¹)₂, —N(R¹)₂,        —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —C(O)N(R¹)₂, R¹, and C₁₋₆ alkyl.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E), R^(A) isindependently selected at each occurrence from:

-   -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —NR¹S(═O)₂R¹,        —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂,        —C(O)N(R¹)₂;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is        independently optionally substituted at each occurrence with one        or more substituents selected from —OR¹, —N(R¹)₂, and 3- to        10-membered heterocycle; and    -   3- to 10-membered heterocycle,    -   wherein each 3- to 10-membered heterocycle in R^(A) is        independently optionally substituted with one or more        substituents selected from —OR¹, —N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹,        —C(O)OR¹, —C(O)N(R¹)₂, and R¹.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E), R^(A) isindependently selected at each occurrence from:

-   -   —N(R¹)₂ and —NR¹C(O)R¹;    -   C₁₋₆ alkyl and —N(R¹)—C₁₋₆ alkyl, each of which is substituted        with 4- to 8-membered heterocycle; and    -   4- to 8-membered heterocycle,    -   wherein each 4- to 8-membered heterocycle in R^(A) is        independently optionally substituted with one or more        substituents selected from —C(O)OR¹ and R¹.

In some embodiments, a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E) comprises aterminal ester of the formula —C(O)OR¹⁵ or —OC(O)R¹⁵, wherein R¹⁵comprises 1 to 12 carbon atoms and at least one basic amine. In someembodiments, the molecular weight of R¹⁵ is between 30 and 200 g/mol.The terminal ester may be metabolizable by one or more hydrolase (e.g.,an esterase) present in human plasma and/or the human liver into acarboxylic acid and an alcohol. The biological activity of the compoundmay be greater than that of the carboxylic acid and the alcohol. Forexample, the compound may exhibit a BEAS2B pIC₅₀ of at least 1 unit orgreater than the carboxylic acid and the alcohol (assessed according tothe assay provided in Example 46). In some embodiments, R¹⁵ is —(C₀₋₄alkyl)(4- to 10-membered heterocycloalkyl), wherein the heterocycloalkylcomprises 1, 2 or 3 nitrogen atoms, and wherein the heterocycloalkyl isoptionally substituted with one or more substituents selected from C₁₋₄alkyl.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E)

-   -   R^(A) is selected from

wherein

-   -   D and E are each independently selected from 3- to 8-membered        heterocycle, each of which is independently optionally        substituted with one or more substituents selected from —NH₂,        —NHCH₃, —N(CH₃)₂, —CH₂CH₂N(CH₃)₂, —C(O)CH₃, —C(O)OH, —C(O)NH₂,        ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, C₃₋₈ carbocycle, and 3- to 6-membered        heterocycle. In some embodiments, D and E are each independently        selected from 4- to 6-membered heterocycle, each of which is        independently optionally substituted with one or more —CH₃. In        some embodiments, D and E are each unsubstituted.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E), R^(A) is—N(R¹)₂, such as

In some embodiments, R^(A) is —NR¹C(O)R¹, such as

In some embodiments, R^(A) is C₁₋₆ alkyl substituted with 4- to8-membered heterocycle wherein the 4- to 8-membered heterocycle issubstituted with —C(O)OR¹, such as

In some embodiments, R^(A) is —N(R¹)—C₁₋₆ alkyl substituted with 4- to8-membered heterocycle, wherein the 4- to 8-membered heterocycle isoptionally substituted, such as

In some embodiments, R^(A) is 4- to 8-membered heterocycle, wherein the4- to 8-membered heterocycle is optionally substituted with one or moresubstituents selected from —C(O)OR¹ and R¹, such as

In some embodiments, R^(A) is —C(O)OR¹, such as

In some embodiments, R^(A) is selected from

In some embodiments, R^(A) is selected from

In some embodiments, R^(A) is selected from

In some embodiments, R^(A) is selected from

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E), R¹ isindependently selected at each occurrence from hydrogen; and C₁₋₆ alkyl,1- to 6-membered heteroalkyl, C₀₋₃ alkyl-(C₃₋₈ carbocycle), and C₀₋₃alkyl-(3- to 10-membered heterocycle), each of which is optionallysubstituted by one or more substituents selected from —NH₂, —NHCH₃,—N(CH₃)₂, —CH₂CH₂N(CH₃)₂, —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH,—CH₂CH₂OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₈carbocycle, and 3- to 6-membered heterocycle. In some embodiments, R¹ isindependently selected at each occurrence from hydrogen; and C₁₋₆ alkyland C₀₋₃ alkyl-(3- to 10-membered heterocycle), each of which isoptionally substituted by one or more substituents selected from —OH,—CH₂OH, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-memberedheterocycle.

In some embodiments, for a compound of Formula (I), (I′), (I-B), (I′-B),(I-C) or (I′-C), B is selected from C₆ 10 aryl and 5- to 10-memberedheteroaryl, such as B is selected from phenyl and pyridyl. In someembodiments, B is pyridyl. In some embodiments, B is 2-pyridyl. In someembodiments, B—(R^(B))_(n) is 2-methylpyridyl.

In some embodiments, for a compound of Formula (I), (I′), (I-B), (I′-B),(I-C) or (I′-C), n is 1 or 2, such as n is 1. In some embodiments, B ispyridyl, n is 1 and R^(B) is in the 2-position.

In some embodiments, for a compound of Formula (I), (I′), (I-B), (I′-B),(I-C) or (I′-C), R^(B) is independently selected at each occurrence fromhalogen, —NH₂, —NHCH₃, —NHCH₂CH₃, —C(O)CH₃, —OH, —OCH₃, —OCH₂CH₃, —CH₃,—CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂F, —CHF₂, and —CF₃. In someembodiments, R^(B) is independently selected at each occurrence fromhalogen, —OH, —CH₃, and —CHF₂. In some embodiments, R^(B) is —CH₃.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E):

-   -   A is selected from phenyl and 5- to 6-membered heteroaryl;    -   B is pyridyl;    -   R^(A) is independently selected at each occurrence from:        -   halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂,            —S(═O)₂NR²R³, —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹,            —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂,            —C(O)NR²R³;        -   C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is            independently optionally substituted at each occurrence with            one or more substituents selected from —OR¹, —N(R¹)₂,            —C(O)OR¹, C₃₋₈ carbocycle, and 3- to 10-membered            heterocycle; and        -   C₃₋₈ carbocycle and 3- to 10-membered heterocycle,        -   wherein each C₃₋₈ carbocycle and 3- to 10-membered            heterocycle in R^(A) is independently optionally substituted            with one or more substituents selected from —OR¹,            —CH₂N(R¹)₂, —N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹,            —C(O)N(R¹)₂, R¹, and C₁₋₆ alkyl;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl, 1- to 6-membered heteroalkyl, C₀₋₃        alkyl-(C_(3-s) carbocycle), and C₀₋₃ alkyl-(3- to 10-membered        heterocycle), each of which is optionally substituted by one or        more substituents selected from —NH₂, —NHCH₃, —N(CH₃)₂,        —CH₂CH₂N(CH₃)₂, —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH,        —CH₂CH₂OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃,        C₃₋₈ carbocycle, and 3- to 6-membered heterocycle;    -   m is 1 or 2;    -   R^(B) is selected from halogen, —OH, —CH₃, and —CHF₂; and    -   n is 1.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E):

-   -   A is phenyl;    -   B is pyridyl;    -   R^(A) is independently selected at each occurrence from:        -   —N(R¹)₂ and —NR¹C(O)R¹;        -   C₁₋₆ alkyl and —N(R¹)—C₁₋₆ alkyl, each of which is            substituted with 4- to 8-membered heterocycle; and        -   4- to 8-membered heterocycle,        -   wherein each 4- to 8-membered heterocycle in R^(A) is            independently optionally substituted with one or more            substituents selected from —C(O)OR¹ and R¹;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl and C₀₋₃ alkyl-(3- to 10-membered heterocycle),        each of which is optionally substituted by one or more        substituents selected from —OH, —CH₂OH, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-membered heterocycle;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E):

-   -   A is phenyl;    -   B is pyridyl;    -   R^(A) is C₁₋₆ alkyl substituted with 4- to 8-membered        heterocycle, wherein the 4- to 8-membered heterocycle is        substituted with —C(O)OR¹;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl and C₀₋₃ alkyl-(3- to 10-membered heterocycle),        each of which is optionally substituted by one or more        substituents selected from —OH, —CH₂OH, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-membered heterocycle;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E):

-   -   A is selected from phenyl and 5- to 6-membered heteroaryl;    -   B is pyridyl;    -   R^(A) is selected from:        -   —C(O)OR¹; and        -   C₀₋₆ alkyl-(3- to 8-membered heterocycle), wherein the 3- to            8-membered heterocycle is substituted with —C(O)OR¹;    -   R¹ is C₀₋₃ alkyl-(3- to 8-membered heterocycle), optionally        substituted by one or more substituents selected from —CH₃,        —CH₂CH₃, —CH(CH₃)₂, and —C(CH₃)₃;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In some embodiments, for a compound of Formula (I):

-   -   A is pyridyl;    -   B is pyridyl;    -   R^(A) is —C(O)OR¹;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl and C₀₋₃ alkyl-(3- to 10-membered heterocycle),        each of which is optionally substituted by one or more        substituents selected from —OH, —CH₂OH, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-membered heterocycle;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In some embodiments, for a compound of Formula (I):

-   -   A is thiophenyl;    -   B is pyridyl;    -   R^(A) is —C(O)OR¹;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₁₋₆ alkyl and C₀₋₃ alkyl-(3- to 10-membered heterocycle),        each of which is optionally substituted by one or more        substituents selected from —OH, —CH₂OH, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, and 3- to 6-membered heterocycle;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In some embodiments, for a compound of Formula (I):

-   -   A is selected from phenyl and thiophenyl;    -   B is pyridyl;    -   R^(A) is —NR¹C(O)R¹;    -   R¹ is independently selected at each occurrence from hydrogen;        and C₀₋₃ alkyl-(3- to 8-membered heterocycle), each of which is        optionally substituted by one or more substituents selected from        —OH, —CH₂OH, —CH₃, —CH₂CH₃, —CH(CH₃)₂, and —C(CH₃)₃;    -   m is 1;    -   R^(B) is —CH₃; and    -   n is 1.

In some embodiments, for a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E):

-   -   A is selected from phenyl and 5- to 6-membered heteroaryl;    -   B is selected from phenyl and pyridyl;    -   R^(A) is selected from

-   -   D and E are each independently selected from 3- to 8-membered        heterocycle, each of which is independently optionally        substituted with one or more substituents selected from —NH₂,        —NHCH₃, —N(CH₃)₂, —CH₂CH₂N(CH₃)₂, —C(O)CH₃, —C(O)OH, —C(O)NH₂,        ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃,        —CH(CH₃)₂, —C(CH₃)₃, C₃₋₈ carbocycle, and 3- to 6-membered        heterocycle;    -   m is 1;    -   R^(B) is selected from halogen, —OH, —CH₃, and —CHF₂; and    -   n is 1.

In some embodiments R^(A) is selected from

In some embodiments, D and E are each independently selected from 4- to6-membered heterocycle, each of which is independently optionallysubstituted with one or more —CH₃. In some embodiments, D and E are eachunsubstituted. In some embodiments, A is selected from phenyl,pyridinyl, thiazolyl and thiophenyl. In some embodiments, A is phenyl.In some embodiments, A is pyridinyl. In some embodiments, A isthiazolyl. In some embodiments, A is thiophenyl.

In some embodiments, a compound of Formula (I), (I′), (I-A), (I′-A),(I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E) is providedas a substantially pure stereoisomer. In some embodiments, thestereoisomer is provided in at least 80% enantiomeric excess, such as atleast 85%, at least 90%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99% or at least 99.9% enantiomeric excess.

In some embodiments, the present disclosure provides a soft ALK5inhibitor. As used herein, the term “soft drug” or “soft ALK5 inhibitor”refers to a biologically active compound that is converted upon enteringthe systemic circulation into a predictable metabolite that exhibitsreduced biological activity relative to the parent compound. A soft drugpreferably exerts its desired therapeutic effect locally at the targetorgan or tissue, then is rapidly converted to a predictable metabolitedesigned to be less active than the parent soft drug upon entering thesystemic circulation, thus reducing systemic exposure to thebiologically active compound. Accordingly, soft drugs have a lowerpotential for undesired side effects relative to non-soft drug compoundshaving comparable biological activity. Preferably, a soft drug of thepresent disclosure exhibits good stability at the intended site ofaction (e.g., the lung), is rapidly metabolized upon entering systemiccirculation, and displays more functional activity than thecorresponding metabolite.

In some embodiments, a soft drug provided herein exhibits an ALK5 pK_(i)of greater than or equal to 9, while the corresponding soft drugmetabolite exhibits an ALK5 pK_(i) of 9 or less, such as 8 or less(assessed according to the assay provided in Example 45). In someembodiments, the difference in pK_(i) of the soft drug and thecorresponding soft drug metabolite is at least 0.5, such as at least0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, orat least 2.0. In some embodiments, a soft drug provided herein exhibitsa BEAS2B pIC₅₀ of greater than or equal to 7, while the correspondingsoft drug metabolite exhibits a BEAS2B pIC₅₀ of 6 or less (assessedaccording to the assay provided in Example 46). In some embodiments, thedifference in pIC₅₀ of the soft drug and the corresponding soft drugmetabolite is at least 1.0, such as at least 1.1, 1.2, 1.3, 1.4, 1.5,1.6, 1.7, 1.8, 1.9, or at least 2.0. In some embodiments, the soft drugand corresponding soft drug metabolite exhibit similar ALK5 pK_(i)values, but the soft drug is more active in cells (e.g., the soft drugexhibits a BEAS2B pIC₅₀ of at least 1.0 greater than the soft drugmetabolite).

In some embodiments, the present disclosure provides a soft ALK5inhibitor comprising an ester. Preferably, the ester inhibits ALK5activity, while the corresponding carboxylic acid of the ester exhibitsreduced ALK5 inhibitory activity. For example, the difference in ALK5pK_(i) of the ester and corresponding acid may be at least 1.0. In someembodiments, a soft drug ester of the present disclosure is administeredto the lung, for example, by inhalation, and inhibits the activity ofALK5 in the lung. However, upon exiting the lung, the ester may bereadily hydrolyzed to the corresponding carboxylic acid, thus reducingsystemic exposure to the ester.

In some embodiments, the present disclosure provides a conjugatecomprising a compound disclosed herein linked, e.g., covalently linked,either directly or through a linker to an antibody construct ortargeting moiety, thereby forming a conjugate. The linker may be anon-cleavable linker or a cleavable linker. A conjugate may berepresented by the formula:

A′

L′-D′)_(p),

wherein A′ is an antibody construct or targeting moiety; L¹ is a linker;D′ is a compound or salt disclosed herein, such as a compound of Formula(I), (I′), (I-A), (I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D),(I-E) or (I′-E); and p is an integer from 1 to 20. In some embodiments,p is an integer from 1 to 10, such as from 1 to 8, 2 to 8, 1 to 6, 3 to5, or from 1 to 3.

In some embodiments, a conjugate is represented by the formula:

A′

D′)_(p),

wherein A′ is an antibody construct or targeting moiety; D′ is acompound or salt disclosed herein, such as a compound of Formula (I),(I′), (I-A), (I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E)or (I′-E); and p is an integer from 1 to 20. In some embodiments, p isan integer from 1 to 10, such as from 1 to 8, 2 to 8, 1 to 6, 3 to 5, orfrom 1 to 3.

Accordingly, a compound or salt of the present disclosure, such as acompound of Formula (I), (I′), (I-A), (I′-A), (I-B), (I′-B), (I-C),(I′-C), (I-D), (I′-D), (I-E) or (I′-E), may be attached to A′ via alinker, L¹, or directly attached to A′ without an intermediate linker.In some embodiments, the compound or salt is covalently attached to anA′ or L¹. It will be understood by the skilled person that not allcompounds of the present disclosure are meant to be attached to L¹ orA′, only those that have suitable attachment sites. A compound or saltdisclosed herein that does not have a suitable attachment site may bemodified to introduce an attachment site.

In some embodiments, L¹ or D′ is bound to A′ via a terminus of an aminoacid sequence or via a side chain of an amino acid, such as the sidechain of lysine, serine, threonine, cysteine, tyrosine, aspartic acid,glutamine, a non-natural amino acid residue, or glutamic acid residue.In some embodiments, L¹ or D′ is bound to A′ via one or more glycans orshort peptide tags of four to six amino acids. L¹ or D′ may beconjugated to A′ via any suitable functional group, such as a thiol, anamine, an amide, an alcohol, a ketone, a carboxylic acid, or an ester.

A linker may be attached to a compound or salt of the present disclosureat any available position. For example, a compound of Formula (I) maycomprise a linker L¹ to A′ in place of R^(A) or through substituentR^(A):

Compounds are typically depicted herein in their unconjugated form,though it will be understood by the skilled person that linker L¹ may becovalently bound to any suitably atom for attachment, such as asubstitutable nitrogen, carbon, sulfur, phosphorous or oxygen of acompound. L¹ may be a cleavable or non-cleavable linker. The linker mayfurther be bound to A′. Preferably, L¹ does not affect the binding ofthe active portions of the conjugate to the binding target(s). Covalentlinkages may be formed by reaction between a functional group on thelinker with a functional group on the compound, and by reaction betweena functional group on the linker with a functional group on A′. As usedherein in the context of conjugates, the term “linker” includes (i)unattached forms of the linker comprising a functional group capable ofcovalently attaching the linker to a compound disclosed herein and afunctional group capable of covalently attaching the linker to anantibody construct or targeting moiety; (ii) partially attached forms ofthe linker bound to a compound disclosed herein, wherein the linkercomprises a functional group capable of covalently attaching the linkerto an antibody construct or targeting moiety; (iii) partially attachedforms of the linker bound to an antibody construct or targeting moiety,wherein the linker comprises a functional group capable of covalentlyattaching the linker to a compound disclosed herein; and (iv) fullyattached forms of the linker bound to both an antibody construct ortargeting moiety and a compound disclosed herein.

Linker L¹ may be short, flexible, rigid, cleavable (e.g., by a lysosomalenzyme), non-cleavable, hydrophilic, or hydrophobic. A linker maycontain segments having different characteristics, such as flexiblesegments and rigid segments. A linker may be chemically stable toextracellular environments, for example, in the bloodstream, or mayinclude moieties that are not stable or are selectively stable. In someembodiments, a linker comprises a moiety that is selectively cleaved,for example, selectively cleaved in cells, a particular organ, or inplasma. A linker may be sensitive to enzymes, such as proteases. Alinker may be insensitive to intracellular processes or proteases. Alinker may be acid-labile, protease-sensitive or photolabile. In someembodiments, a linker comprises a peptide, succinimide, maleimide,polyethylene glycol, alkylene, alkenylene, alkynylene, disulfide,hydrazone, polyether, polyester, polyamide, aminobenzyl-carbamate, or acombination thereof.

In some aspects, the present disclosure provides a compound of Formula(I), (I′), (I-A), (I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D),(I-E) or (I′-E), or a compound disclosed in Table 1 or Table 2, whereinthe compound is covalently bound to A′, optionally via linker L¹. Insome embodiments, the antibody construct is an antibody. In someembodiments, the present disclosure provides a compound of Formula (I),(I′), (I-A), (I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E)or (I′-E), or a compound disclosed in Table 1 or Table 2, wherein thecompound is covalently bound to a linker, L¹, to form a compound-linker.A′ or L¹ may be covalently attached to any position of the compound,valence permitting. A linker L¹ disclosed herein may comprise from about10 to about 500 atoms, such as 10 to 400 atoms, 10 to 300 atoms, 30 to400 atoms, or 30 to 300 atoms.

The targets of the antibody, antibody construct, or targeting moiety maydepend on the desired therapeutic applications of the conjugate.Typically, the targets are molecules present on the surfaces of cellsinto which it is desirable to deliver an ALK5 inhibitor, such as Tcells, and the antibodies preferably internalize upon binding to thetarget. For applications in which the conjugates are intended tostimulate the immune system by reducing TGF-β activity, it may bedesirable to generate antibodies, antibody constructs, or targetingmoieties that bind to T cell surface molecules. Not wishing to be boundby any particular theory, it is believed that the delivery of ALK5inhibitors to T cells can activate CD4⁺ and/or CD8⁺ T cell activity andinhibit regulatory T cell activity, both of which contribute to immunetolerance of tumors. Accordingly, antibodies, antibody constructs, ortargeting moieties (A′) that bind to T cell surface molecules in theconjugates of the present disclosure are useful for the treatment ofvarious cancers, such as those described herein below. In someembodiments, A′ binds to CD4⁺ T cells, CD8⁺ T cells, T_(REG) cells, orany combination thereof. In some embodiments, A′ binds to a pan T cellsurface molecule, such as CD1, CD2, CD3, CD4, CD5, CD6, CD7, CD8, CD25,CD28, CD70, CD71, CD103, CD184, Tim3, LAG3, CTLA4, or PD1. Examples ofantibodies that bind to T cell surface molecules and are believed to beinternalizing include OKT6, OKT11, OKT3, OKT4, OKT8, 7D4, OKT9, CD28.2,UCHT1, M290, FR70, pembrolizumab, nivolumab, cemiplimab, anddostarlimab.

An antibody, antibody construct, or targeting moiety disclosed hereinmay comprise an antigen binding domain that specifically binds to atumor antigen or antigen associated with the pathogenesis of fibrosis.In some embodiments, the antigen binding domain specifically binds to anantigen on a T cell, a B cell, a stellate cell, an endothelial cell, atumor cell, an APC, a fibroblast cell, a fibrocyte cell, or a cellassociated with the pathogenesis of fibrosis. In some embodiments, theantigen binding domain targets CTLA4, PD-1, OX40, LAG-3, GITR, GARP,CD25, CD27, PD-L1, TNFR2, ICOS, 41BB, CD70, CD73, CD38 or VTCN1. In someembodiments, the antigen binding domain targets PDGFRβ, integrin αvβ1,integrin αvβ3, integrin αvβ6, αvβ8, endosialin, FAP, ADAM12, LRRC15,MMP14, PDPN, CDH11, F2RL2, ASGR1, or ASGR2.

The chemical entities described herein can be synthesized according toone or more illustrative schemes herein and/or techniques known in theart. Materials used herein are either commercially available or preparedby synthetic methods generally known in the art. These schemes are notlimited to the compounds listed in the examples or by any particularsubstituents, which are employed for illustrative purposes. Althoughvarious steps are described and depicted in Schemes 1 and 2 and Examples1-44, the steps in some cases may be performed in a different order thanthe order shown in Schemes 1 and 2 and Examples 1-44. Variousmodifications to these synthetic reaction schemes may be made and willbe suggested to one skilled in the art having referred to the presentdisclosure. Numberings or R groups in each scheme typically have thesame meanings as those defined elsewhere herein unless otherwiseindicated.

Unless specified to the contrary, the reactions described herein takeplace at atmospheric pressure, generally within a temperature range from−10° C. to 200° C. Further, except as otherwise specified, reactiontimes and conditions are intended to be approximate, e.g. taking placeat about atmospheric pressure within a temperature range of about −10°C. to about 110° C. over a period of about 1 to about 24 hours;reactions left to run overnight average a period of about 16 hours.

In general, compounds of the disclosure may be prepared by the followingreaction schemes:

In some embodiments, a compound of Formula 1i may be prepared accordingto Scheme 1. For example, imidate 1b can be formed from nitrile 1a via aPinner Reaction, then converted to amidine 1c in the presence ofammonium chloride. Cyclization with methyl 3-methoxyacrylate (1d) mayproceed to give pyrimidinone 1e, which may be treated with phosphorusoxychloride to give chloropyrimidine 1f. Chloropyrimidine 1f can besubjected to a C—N coupling reaction—optionally a Pd-catalyzed couplingreaction such as a Buchwald-Hartwig amination—with2-chloropyrimidin-4-amine (1g) to provide heteroaryl amine 1h.Optionally, 1h may be subjected to one or more coupling reactions, andoptionally one or more protecting group manipulations, to provide acompound of Formula 1i.

In some embodiments, a compound of Formula 2f may be prepared accordingto Scheme 2. For example, nitro compound 2a can be reduced to amine 2b.The amine may then be subjected to a C—N coupling reaction-optionally aPd-catalyzed coupling reaction such as a Buchwald-Hartwig amination—withbis(2-chloropyrimidin-4-yl)amine (2c) to provide heteroaryl amine 2d. Asubstitution reaction can provide heteroaryl iodide 2e, which mayundergo a nucleophilic aromatic substitution reaction, and optionallyone or more protecting group manipulations, to provide a compound ofFormula 2f.

In some embodiments, a compound of the present disclosure, for example,a compound of a formula given in Table 1 or 2, is synthesized accordingto one of the general routes outlined in Schemes 1 and 2, Examples 1-44,or by methods generally known in the art. In some embodiments, exemplarycompounds may include, but are not limited to, a compound or saltthereof selected from Table 1.

TABLE 1 [M + No. Structure Chemical Name H]⁺ 1

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [[(3S)-pyrrolidin-3-yl]amino]phenyl]pyrimidine-2,4- diamine 440.2 2

[2-chloro-6-fluoro-4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]- piperazin-1-yl- methanone 520.13

1-[3-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]anilino] methyl]azetidin- 1-yl]ethanone 482.3 4

N2-[4-[4-(4-methylpiperazin-1- yl)-1-piperidyl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 537.2 5

N2-(4-aminophenyl)-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 371.1 6

N-[2-methoxy-5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 512.2 7

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[rac-(3R)-3-methylpiperazin-1- yl]phenyl]pyrimidine-2,4- diamine 454.1 8

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4- thiomorpholinophenyl)pyrimidine-2,4-diamine 457.2 9

[(1R)-1-methylpropyl] 2-[1-[4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 554.5 10

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperidine-3- carboxamide 482.1 11

pyrrolidin-3-yl 1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-3- carboxylate 552.3 12

1-[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl] sulfonylpiperazin- 1-yl]ethanone 546.0 13

[(1R)-1-methylpropyl] 2-[5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetate 554.3 14

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperidine-4- carboxamide 482.1 15

N2-[4-(2,6- diazaspiro[3.3]heptan-2- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 452.2 16

3-[4-[[2-[4-[4-(dimethylamino)- 1-piperidyl]anilino]pyrimidin-4-yl]amino]pyrimidin-2-yl]-4- methyl-phenol 497.2 17

azetidin-3-yl 1-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-4-carboxylate552.1 18

N-[2-methoxy-4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl] methanesulfonamide 479.0 19

N4-[2-(5-chloro-2,4-difluoro- phenyl)pyrimidin-4-yl]-N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 496.1 20

N-[2-methoxy-5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 3-pyridyl]methanesulfonamide 480.0 21

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4- piperazin-1-ylsulfonylphenyl)pyrimidine- 2,4-diamine 504.2 22

4-piperidyl 1-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] azetidine-3-carboxylate552.3 23

N2-[4-[(4-benzylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 544.2 24

N2-[4- [(dimethylamino)methyl]phenyl]- N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 413.2 25

methyl 1-[[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylate512.2 26

4-[[2-(4-aminoanilino)pyrimidin- 4-yl]amino]-2-(6-methyl-2-pyridyl)pyrimidine-5-carboxylic acid 415.0 27

N2-[4-(4-benzylpiperazin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 530.1 28

methyl 1-[[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-4-carboxylate511.1 29

(1-methylpyrrolidin-3-yl) 1-[[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 580.2 30

[2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazol-5-yl]-piperazin- 1-yl-methanone 475.1 31

2-[1-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazin-2- yl]acetic acid 498.4 32

N2-[4-[[(2R,5R)-2,5- dimethylpiperazin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 33

azetidin-3-ylmethyl 4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 469.1 34

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [rac-(1S,5R)-3,8-diazabicyclo[3.2.1]octan-3- yl]phenyl]pyrimidine-2,4- diamine 466.2 35

N4-[2-(2,4- difluorophenyl)pyrimidin-4-yl]- N2-(4- morpholinophenyl)pyrimidine-2,4-diamine 462.1 36

2-[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]pyrazol-1- yl]ethanol 466.1 37

4-hydroxy-2-[2-[4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-1-yl]-2-oxo-ethyl]sulfanyl-butanoic acid 616.3 38

N2-[4-(4-ethylpiperazin-1- yl)sulfonylphenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 532.1 39

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[3-(thiomorpholinomethyl)phenyl] pyrimidine-2,4-diamine 471.1 40

N-[3-(hydroxymethyl)pyrrolidin- 3-yl]-4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxamide 504.2 41

isobutyl 2-[1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 554.5 42

N-(azetidin-3-ylmethyl)-2-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4-carboxamide 474.9 43

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-phenyl-pyrimidine-2,4-diamine 356.2 44

N-(azetidin-3-yl)-2-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4- carboxamide 461.1 45

[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methanol 386.2 46

[(2S)-4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]methanol 470.1 47

ethyl 2-[4-[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]pyrazol-1- yl]acetate 508.1 48

methyl 6-[[2-[4-(4- methylpiperazin-1- yl)anilino]pyrimidin-4-yl]amino]-2-(6-methyl-2- pyridyl)pyrimidine-4-carboxylate 512.2 49

N2-[4-chloro-3-(piperazin-1- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 488.2 50

azetidin-3-yl 1-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] pyrrolidine-3-carboxylate538.1 51

[(3S)-1-methylpyrrolidin-3-yl] 4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]benzoate 483.0 52

[(3R)-1-methylpyrrolidin-3-yl] 5-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 489.1 53

N2-[1-(2-aminoethyl)pyrazol-4- yl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 389.2 54

2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenoxy]ethanol 416.1 55

(1-methylpyrrolidin-3-yl) 1-[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-3-carboxylate 566.3 56

N2-[4-(1-methylpyrazol-4- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 436.1 57

azetidin-3-ylmethyl 2-[5-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetate 567.1 58

3-hydroxy-N-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]pyrrolidine-3- carboxamide 484.159

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl] methyl]-N-[rac- (3S)-pyrrolidin-3- yl]piperidine-3-carboxamide 565.2 60

methyl 1-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxylate 497.1 61

[2-methyl-5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methanol 400.3 62

methyl 4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]benzoate 414.0 63

N-(2-aminoethyl)-N-methyl-4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzenesulfonamide 492.2 64

N2-[4-[[(3R)-3- (methoxymethyl)piperazin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 498.1 65

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [[(2R)-pyrrolidin-2-yl]methylamino]phenyl] pyrimidine-2,4-diamine 454.2 66

4-piperidyl 3-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 483.2 67

azetidin-3-ylmethyl (3S)-1-[[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 566.1 68

N4-[2-(5-chloro-2-fluoro- phenyl)-5-cyclopropyl-pyrimidin-4-yl]-N2-(4-piperazin- 1-ylphenyl)pyrimidine-2,4- diamine517.1 69

[(3R)-1-methylpyrrolidin-3-yl] 1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]azetidine-3-carboxylate 552.1 70

N-[2-(dimethylamino)ethyl]-4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzenesulfonamide 506.1 71

1-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperidine-4- carboxylic acid 482.2 72

[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-morpholino- methanone 469.2 73

(1-methylazetidin-3-yl)methyl (3S)-1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 580.2 74

(3S)-1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] pyrrolidin-3-ol 455.1 75

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [[rac-(1S,5R)-3,8-diazabicyclo[3.2.1]octan-3- yl]methyl]phenyl]pyrimidine- 2,4-diamine480.2 76

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]azetidine-3- carboxamide 454.1 77

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-1- tetrahydropyran-4-yl-piperidine- 4-carboxamide 566.278

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]pyrrolidine-3- carboxamide 468.0 79

N2-(3-methoxy-4-morpholino- phenyl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 471.2 80

N2-[4-(4-ethylpiperazin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.2 81

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-methyl-3-[[rac-(3R,5S)-3,5- dimethylpiperazin-1-yl]methyl]phenyl]pyrimidine- 2,4-diamine 496.1 82

2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]acetic acid 414.1 83

methyl 1-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-3- carboxylate 497.1 84

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[1-(2-morpholinoethyl)pyrazol-4- yl]pyrimidine-2,4-diamine 459.2 85

4-piperidyl 5-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-3- carboxylate 489.1 86

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(2-piperazin-1-ylpyrimidin-5- yl)pyrimidine-2,4-diamine 442.2 87

N-methyl-3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl] amino]-N-(4- piperidyl)benzenesulfonamide 532.288

azetidin-3-ylmethyl 1-[[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] azetidine-3-carboxylate538.2 89

N-(azetidin-3-ylmethyl)-4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxamide 474.0 90

N4-[2-(2- fluorophenyl)pyrimidin-4-yl]- N2-[4-[4-(methylamino)-1-piperidyl]phenyl]pyrimidine- 2,4-diamine 471.1 91

N-ethyl-2-[2-(6-methyl-2- pyridyl)-6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4- yl]acetamide 525.1 92

N-(azetidin-3-ylmethyl)-4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzamide 468.1 93

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazine- 1-sulfonamide 519.1 94

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[5-(piperazin-1-ylmethyl)-3- thienyl]pyrimidine-2,4-diamine 460.2 95

2-[5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetic acid498.4 96

azetidin-3-ylmethyl 4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 475.1 97

[(3S)-1-methylpyrrolidin-3-yl] 1- [[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]azetidine-3-carboxylate 552.1 98

N2-[4-(3-aminoazetidin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 426.1 99

[3-methyl-1-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] azetidin-3-yl]methanol469.2 100

azetidin-3-ylmethyl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 475.0 101

pyrrolidin-3-yl 1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxylate 552.2102

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [4-(pyrrolidin-3-ylmethyl)piperazin-1- yl]phenyl]pyrimidine-2,4- diamine 523.1 103

[rac-(3R)-1-methylpyrrolidin-3- yl] 2-[1-[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2-yl]acetate 581.1 104

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(thiomorpholinomethyl)phenyl] pyrimidine-2,4-diamine 471.1 105

[rac-(3R)-pyrrolidin-3-yl] 1-[[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 566.2 106

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-1-(3- piperidyl)methanesulfonamide 532.1 107

N-isopropyl-4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]sulfonyl- piperazine-1-carboxamide589.1 108

4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]benzenesulfonamide 435.0 109

N2-[4-[(4-tert-butylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 510.3 110

4-isopropyl-N-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-1- carboxamide 525.2111

N2-[4-fluoro-3-(piperazin-1- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 472.2 112

[1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl]-3- piperidyl]-piperazin-1-yl- methanone 565.1113

[(1S)-1-methylpropyl] 4-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxylate 540.2114

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[5- (4-piperidyl)-2-pyridyl]pyrimidine-2,4-diamine 440.3 115

[5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-thienyl]-piperazin-1-yl- methanone474.1 116

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[methyl-[(3S)-pyrrolidin-3- yl]amino]phenyl]pyrimidine-2,4- diamine454.3 117

N-isobutyl-2-[2-(6-methyl-2- pyridyl)-6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4- yl]acetamide 553.2 118

N2-[3-(azetidin-3- ylmethylamino)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 440.4 119

6-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-3-sulfonamide 436.0 120

N2-(3-fluoro-4-piperazin-1-yl- phenyl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 458.1 121

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(5- piperazin-1-yl-2-pyridyl)pyrimidine-2,4-diamine 441.3 122

N-methyl-4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- N-(4- piperidyl)benzenesulfonamide 532.2123

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- (4-pyrrolidin-1-yl-1-piperidyl)phenyl]pyrimidine- 2,4-diamine 508.2 124

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] pyrrolidine-3-carboxylic acid 483.0 125

1-(azetidin-3-yl)-3-[3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]urea 469.1 126

N2-(3-amino-4-chloro-phenyl)- N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 405.0 127

N2-[4-[(3-methoxyazetidin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 455.2 128

N-[2-methyl-5-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 496.2129

N2-[4-(3,4,6,7,8,8a-hexahydro- 1H-pyrrolo[1,2-a]pyrazin-2-ylmethyl)phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 494.2 130

N2-[4-fluoro-3-[[rac-(3R,5S)- 3,5-dimethylpiperazin-1-yl]methyl]phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 500.1 131

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] azetidin-3-ol 441.1 132

[(2R)-1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazin-2-yl]methanol484.1 133

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(4-piperidyl)phenyl]pyrimidine- 2,4-diamine 439.2 134

1-[2-(dimethylamino)ethyl]-N- [3-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4-carboxamide 553.3 135

N-[2-chloro-5-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 516.2136

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[4-(2-pyridylmethyl)piperazin-1- yl]phenyl]pyrimidine-2,4- diamine 531.1137

N4-[2-(5-chloro-2-fluoro- phenyl)pyrimidin-4-yl]-N2-[4-[4-(methylamino)-1- piperidyl]phenyl]pyrimidine-2,4- diamine 505.1 138

azetidin-3-yl 1-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxylate552.2 139

[(3S)-1-methylpyrrolidin-3-yl] (3R)-1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 580.2 140

N-[[3-(hydroxymethyl)azetidin- 3-yl]methyl]-2-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiazole-4-carboxamide 505.1 141

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[3-(morpholinomethyl)phenyl] pyrimidine-2,4-diamine 455.1 142

azetidin-3-ylmethyl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-3- carboxylate 475.1 143

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [rac-(3S,5R)-3,5-dimethylpiperazin-1- yl]phenyl]pyrimidine-2,4- diamine 468.2 144

[(3S)-pyrrolidin-3-yl] 1-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine- 3-carboxylate 552.3145

1-methyl-N-[3-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 496.2146

2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]ethanol 400.2 147

4-piperidyl 4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 483.1 148

4-piperidyl 2-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4-carboxylate 490.0 149

3-[2-[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-1-yl]- 2-oxo-ethyl]sulfanyltetrahydrofuran-2- one 598.3 150

1-methyl-N-[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 496.2151

N2-[4-(4-aminocyclohexen-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 451.2 152

[(3R)-pyrrolidin-3-yl] 1-[[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] azetidine-3-carboxylate538.1 153

N2-[4-(4-methylpiperazin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 454.2 154

N-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]azetidine-3- carboxamide 454.1 155

azetidin-3-ylmethyl (3R)-1-[[2- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiazol-4-yl]methyl]piperidine- 3-carboxylate 573.1 156

ethyl 2-[5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetate 526.3157

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- (2-piperazin-1-ylethylamino)phenyl]pyrimidine- 2,4-diamine 483.2 158

N-[2-chloro-4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 516.0159

azetidin-3-yl 2-[1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 553.1 160

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2- (2,3,4,5-tetrahydro-1H-3-benzazepin-7-yl)pyrimidine-2,4- diamine 425.1 161

[(3S)-1-methylpyrrolidin-3-yl] 1- [[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]pyrrolidine-3-carboxylate 566.1 162

4-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]sulfamoyl] benzamide 554.2 163

N2-[4-(2,6- diazaspiro[3.3]heptan-2- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 466.2 164

3-(hydroxymethyl)-N-[3-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]azetidine-3- carboxamide 484.0 165

N4-[2-(5-chloro-2-fluoro- phenyl)pyrimidin-4-yl]-N2-[4-(morpholinomethyl)phenyl] pyrimidine-2,4-diamine 492.1 166

4-methyl-3-[4-[[2-(4- morpholinoanilino)pyrimidin-4-yl]amino]pyrimidin-2-yl]phenol 456.1 167

azetidin-3-yl 1-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] azetidine-3-carboxylate524.2 168

1-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-3-pyrrolidin-3- yl-urea 483.1 169

N2-[4-[1-(2- methoxyethyl)pyrazol-4- yl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 480.0 170

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(p-tolyl)pyrimidine-2,4-diamine 370.1 171

methyl 1-isopropyl-4-[2-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]anilino]ethyl] piperazine-2-carboxylate583.1 172

methyl (2R)-4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxylate 498.0173

azetidin-3-yl 1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-3- carboxylate 538.3174

[(3S)-pyrrolidin-3-yl] 1-[[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] azetidine-3-carboxylate538.2 175

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(1,2,3,4-tetrahydroisoquinolin-7- yl)pyrimidine-2,4-diamine 411.1 176

N2-[3-[[4-(methylamino)-1- piperidyl]sulfonyl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 532.2 177

N2-[4-(3,9- diazabicyclo[3.3.1]nonan-3- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 494.3 178

azetidin-3-ylmethyl 3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 469.1 179

4-methyl-3-[4-[[2-(4-piperazin- 1-ylanilino)pyrimidin-4-yl]amino]pyrimidin-2-yl]phenol 455.1 180

3-hydroxy-N-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]azetidine-3- carboxamide 470.0 181

N2-[1-(azetidin-3- ylmethyl)pyrazol-4-yl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 415.1 182

(1-methylazetidin-3-yl)methyl (3R)-1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 580.1 183

[(2R)-4-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazin-2-yl]methanol484.1 184

N-[2-chloro-6-fluoro-4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 534.1185

[(3S)-pyrrolidin-3-yl] 1-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine- 4-carboxylate 552.1186

N2-[4-[3-(aminomethyl)azetidin- 1-yl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 440.2 187

N2-[4-(aminomethyl)phenyl]- N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 385.2 188

[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-thienyl]-piperazin-1- yl-methanone474.1 189

N2-[4-(4-isopropylpiperazin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.1 190

1-[4-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazin-1- yl]ethanone 482.2 191

N2-(5-amino-2,3-dihydro-1,4- benzodioxin-7-yl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 429.2 192

[rac-(1R)-1-methylpropyl] 4-[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2-carboxylate 540.2 193

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(1,2,3,6-tetrahydropyridin-4- yl)phenyl]pyrimidine-2,4- diamine 437.1194

N2-[4-[4-(isobutylamino)-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 510.3 195

[(3R)-1-methylpyrrolidin-3-yl] 1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]pyrrolidine-3-carboxylate 566.1 196

N2-[4-(1,4-diazepan-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 454.1 197

N2-[4-(3,8- diazabicyclo[3.2.1]octan-8- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 480.2 198

N2-(2-isopropyl-3,4-dihydro-1H- isoquinolin-7-yl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 453.2 199

N2-[4-(azetidin-3- ylamino)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 426.1 200

(2R)-4-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylicacid 498.2 201

methyl 1-methyl-4-[[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylate526.1 202

N2-[4-[azetidin-3- ylmethyl(methyl)amino]phenyl]- N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 454.2 203

3-hydroxy-N-[3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]pyrrolidine-3- carboxamide 484.1204

methyl 1-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxylate 498.3205

isobutyl 2-[5-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetate 554.4206

3-(hydroxymethyl)-N-[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-thienyl]azetidine-3- carboxamide 490.2207

azetidin-3-yl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-3- carboxylate 461.1 208

N-(2-aminoethyl)-4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzenesulfonamide 478.2 209

3-(hydroxymethyl)-N-[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]azetidine-3- carboxamide 484.0 210

[(3S)-1-methylpyrrolidin-3-yl] (3S)-1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 580.2 211

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-thienyl]azetidine-3- carboxamide 460.0212

[rac-(3S)-pyrrolidin-3-yl] 1-[[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 566.2 213

N2-[4-(4-isopropylpiperazin-1- yl)sulfonylphenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 546.1 214

N-(azetidin-3-yl)-1-[[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxamide551.2 215

4-piperidyl 1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-3- carboxylate 566.2216

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[rac-(2S)-2-methylpiperazin-1- yl]phenyl]pyrimidine-2,4- diamine 454.1217

[(3S)-1-methylpyrrolidin-3-yl] 4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 489.1 218

4-piperidyl 1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxylate 566.1219

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [4-[(2-pyridylmethylamino)methyl]-1- piperidyl]phenyl]pyrimidine- 2,4-diamine559.2 220

azetidin-3-yl 1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine- 4-carboxylate 538.1221

N2-[4-(3,8- diazabicyclo[3.2.1]octan-8- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 466.1 222

N4-[6-methyl-2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(4-methylpiperazin-1- yl)phenyl]pyrimidine-2,4- diamine 468.1 223

N2-(3-isopropyl-1,2,4,5- tetrahydro-3-benzazepin-7-yl)-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 467.3224

(3S)-1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxylicacid 497.2 225

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(1,2,3,4-tetrahydroquinolin-7- yl)pyrimidine-2,4-diamine 411.2 226

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4-methylthiazol-2-yl)pyrimidine- 2,4-diamine 377.0 227

N-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]pyrrolidine-3- carboxamide 468.1 228

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazine- 1-carboxamide 483.1 229

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperidine- 4-sulfonamide 518.2 230

[rac-(1R)-1-methylpropyl] 1-[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2-carboxylate 540.2 231

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4- pyrrolidin-3-ylphenyl)pyrimidine-2,4-diamine 425.1 232

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(5,6,7,8-tetrahydroimidazo[1,2- a]pyrazin-2- yl)phenyl]pyrimidine-2,4-diamine 477.2 233

N2-[3-(4-methylpiperazin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 454.2 234

N2-(4-bromophenyl)-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 435.8 235

5-[[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazin-1- yl]methyl]tetrahydrofuran-2-one 538.2 236

methyl 1-methyl-4-[2-[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]anilino]ethyl] piperazine-2-carboxylate555.1 237

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] azetidine-3-carboxylic acid 469.2 238

N2-[4-chloro-3-[(4- methylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 502.1 239

4-piperidyl 5-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 489.1 240

N2-[4-[[(3R,5S)-3,5- dimethylpiperazin-1- yl]methyl]phenyl]-N4-[2-(6methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 241

ethyl 2-[1-[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 526.4 242

N2-[3-fluoro-4- (morpholinomethyl)phenyl]-N4- [2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 473.2 243

N2-[4-[4-[methyl(2- pyridylmethyl)amino]-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 559.3 244

N2-[4-fluoro-3-[(4- methylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 486.2 245

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(1-pyrrolidin-3-ylpyrazol-4- yl)pyrimidine-2,4-diamine 415.1 246

N4-[2-(5-chloro-2,4-difluoro- phenyl)pyrimidin-4-yl]-N2-[4-[4-(dimethylamino)-1- piperidyl]phenyl]pyrimidine-2,4- diamine 538.1 247

N2-[4-methyl-3-(piperazin-1- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.2 248

(1-methylpyrrolidin-3-yl) 1-[[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-4-carboxylate 580.2 249

N2-[4-[(3,3-dimethylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.3 250

N2-[4-(2- methoxyethoxy)phenyl]-N4-[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 430.2 251

N2-[4-[4-(methylamino)-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.1 252

methyl 2-[5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetate 512.4253

N2-[4-[2- (dimethylamino)ethyl]phenyl]- N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 427.2 254

methyl (2S)-4-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylate512.2 255

4-methyl-3-[4-[[2-[4-[4-(4- methylpiperazin-1-yl)-1-piperidyl]anilino]pyrimidin-4- yl]amino]pyrimidin-2-yl]phenol 552.2 256

azetidin-3-ylmethyl 2-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4-carboxylate 476.0 257

[(3S)-pyrrolidin-3-yl] 3-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 469.2 258

azetidin-3-yl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 461.1 259

N2-[4-(2- aminoethylamino)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 414.2 260

azetidin-3-yl 3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 455.1 261

1-[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazin-1- yl]ethanone 482.2 262

N2-[4-[[3-(dimethylamino)-3- methyl-azetidin-1-yl]methyl]phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 482.2 263

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [2-[rac-(3S,5R)-3,5-dimethylpiperazin-1- yl]ethylamino]phenyl] pyrimidine-2,4-diamine 511.2264

N2-[4-(4-tert-butylpiperazin-1- yl)sulfonylphenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 560.1 265

N-[2-methyl-4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine- 4-carboxamide 496.1266

[1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] azetidin-3-yl]methanol 455.1 267

N2-[5-(4-methylpiperazin-1-yl)- 2-pyridyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 455.1 268

methyl (2R)-4-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylate512.2 269

1-[2-(dimethylamino)ethyl]-N- [4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4-carboxamide 553.2 270

N2-[4-(4-methylpiperazin-1- yl)sulfonylphenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 518.2 271

(2S)-N-methyl-4-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxamide511.1 272

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl] methanesulfonamide 449.1 273

N-[2-(methylamino)ethyl]-3-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzenesulfonamide 492.1 274

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[methyl-[rac-(3R)-pyrrolidin-3- yl]amino]phenyl]pyrimidine-2,4- diamine454.1 275

azetidin-3-ylmethyl 1-[[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-4-carboxylate566.1 276

(3R)-3-(hydroxymethyl)-N-[4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine- 1-carboxamide 513.2277

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-spiro[indane-2,3′-pyrrolidine]-5- yl-pyrimidine-2,4-diamine 451.1 278

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[[rac-(3R)-pyrrolidin-3- yl]amino]phenyl]pyrimidine- 2,4-diamine 440.1279

N2-[4-[[(1R,4R)-2,5- diazabicyclo[2.2.1]heptan-2-yl]methyl]phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 466.3 280

N2-[4-(azetidin-3- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 425.3 281

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(5- morpholino-2-pyridyl)pyrimidine-2,4-diamine 442.3 282

1-(1-methyl-4-piperidyl)-N-[3- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 579.2283

(2S)-4-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylicacid 498.2 284

N2-(1H-indazol-5-yl)-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 396.0 285

4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenol 372.2 286

N2-[1-(3-aminopropyl)pyrazol- 4-yl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 403.2 287

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(piperazin-1-ylmethyl)thiazol-2- yl]pyrimidine-2,4-diamine 461.2 288

N-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperidine-4- carboxamide 482.2 289

N2-[4-(3-aminopyrrolidin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 440.2 290

[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-piperazin- 1-yl-methanone 468.1 291

[(3R)-1-methylpyrrolidin-3-yl] 2-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazole-4-carboxylate 490.0 292

(4-isopropylpiperazin-1-yl)-[3- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methanone510.1 293

(2R)-N-methyl-4-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2- carboxamide511.1 294

[2-fluoro-5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methanol 404.2 295

[2-chloro-5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methanol 420.1 296

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]azetidine-3- sulfonamide 490.1 297

[(3S)-1-methylpyrrolidin-3-yl] 1- [[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-4-carboxylate 580.2 298

N-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazine- 1-carboxamide 483.1 299

3-hydroxy-N-[3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]azetidine-3- carboxamide 470.0 300

N2-(3-fluoro-4-morpholino- phenyl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 459.1 301

2-[4-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] piperazin-1-yl]ethanol 498.2 302

N2-[4-[4-(aminomethyl)-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.4 303

4-methyl-N-[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine- 1-sulfonamide 533.1304

ethyl 2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]acetate 442.1 305

N2-[4-[(4-methylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.1 306

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(3- piperazin-1-ylsulfonylphenyl)pyrimidine- 2,4-diamine 504.2 307

methyl (2S)-4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxylate 498.1308

[(3S)-1-methylpyrrolidin-3-yl] 3- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]benzoate 483.1 309

N-methyl-2-[2-(6-methyl-2- pyridyl)-6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4- yl]acetamide 511.2 310

[3-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]anilino]methyl] azetidin-3-yl] methanol 470.2 311

N2-[4-methyl-3-[(4- methylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 312

methyl 5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-benzoate 498.2 313

[(3R)-pyrrolidin-3-yl] 1-[[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-4-carboxylate566.1 314

N4-[2-(5-chloro-2,4-difluoro- phenyl)pyrimidin-4-yl]-N2-[4-(4-pyrrolidin-1-yl-1- piperidyl)phenyl]pyrimidine- 2,4-diamine 563.1 315

N2-(3-aminophenyl)-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 371.2 316

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]acetamide 413.1 317

(4-isopropylpiperazin-1-yl)-[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methanone510.2 318

azetidin-3-ylmethyl 1-[[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxylate566.2 319

(3R)-1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] pyrrolidin-3-ol 455.1 320

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl]-N-(4- piperidyl)piperidine-3- carboxamide 579.3321

2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]anilino]ethanol 415.0 322

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(3-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 440.2 323

[(3S)-1-methylpyrrolidin-3-yl] 2- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazole-4-carboxylate 490.0 324

N2-[4-(4-amino-1- piperidyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 454.1 325

azetidin-3-yl 2-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-5-carboxylate 462.0 326

N4-[2-(5-chloro-2,4-difluoro- phenyl)pyrimidin-4-yl]-N2-[4-[4-(4-methylpiperazin-1-yl)-1- piperidyl]phenyl]pyrimidine-2,4- diamine592.2 327

N-(azetidin-3-yl)-4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxamide 460.1 328

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- (1-methyltriazol-4-yl)phenyl]pyrimidine-2,4- diamine 437.0 329

[(2R)-4-[[2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazol-4- yl]methyl]piperazin-2-yl]methanol 491.1 330

4-methyl-N-[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-1- carboxamide 497.2331

N2-[4-(2,5-dihydro-1H-pyrrol-3- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 423.2 332

N2-[3-(3,8- diazabicyclo[3.2.1]octan-3- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 480.2 333

N2-[4-(1-benzyltriazol-4- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 513.0 334

(2R)-N-methyl-4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxamide 497.1335

N2-[3-methoxy-4-[[rac-(3S,5R)- 3,5-dimethylpiperazin-1-yl]methyl]phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 512.3 336

4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- N-[(3R)-pyrrolidin-3-yl]thiophene-2-carboxamide 474.2 337

[(3R)-pyrrolidin-3-yl] 1-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine- 4-carboxylate 552.2338

azetidin-3-ylmethyl 2-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-5-carboxylate 476.0 339

(3R)-1-[[2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazol-4- yl]methyl]piperidine-3-carboxylic acid 504.2 340

N4-[5-(6-methyl-2-pyridyl)-3H- triazolo[4,5-d]pyrimidin-7-yl]- N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 482.1 341

[(3R)-1-methylpyrrolidin-3-yl] (3R)-1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 580.3 342

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [[(2S)-pyrrolidin-2-yl]methylamino]phenyl] pyrimidine-2,4-diamine 454.2 343

N2-[4-(4-amino-1-piperidyl)-3- fluoro-phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 472.2 344

N2-[4-[[(2S,5R)-2,5- dimethylpiperazin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 345

[(3S)-1-methylpyrrolidin-3-yl] 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-3-carboxylate 489.1 346

N2-[4-[2-(4-isopropylpiperazin- 1-yl)ethylamino]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 525.2 347

N2-[4-[1-[2- (methylamino)ethyl]pyrazol-4- yl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 479.2 348

N2-(2-methoxy-4-morpholino- phenyl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 471.3 349

[(3R)-3- (hydroxymethyl)piperazin-1-yl]- [2-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiazol-4-yl]methanone 505.0 350

N2-[4-chloro-3- (thiomorpholinomethyl)phenyl]- N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 505.2 351

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]- N2-(4-morpholin-3-ylphenyl)pyrimidine- 2,4-diamine 441.1 352

N2-[4-[[4-(methylamino)-1- piperidyl]sulfonyl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 532.2 353

N-(azetidin-3-ylmethyl)-N-[4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]acetamide 482.3 354

N4-[2-(5-fluoro-6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 459.2 355

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]- N2-[4-(2-morpholinoethylamino)phenyl] pyrimidine-2,4-diamine 484.1 356

N2-[4-[1-[2- (dimethylamino)ethyl]pyrazol-4-yl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 493.1 357

N-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-1- tetrahydropyran-4-yl-piperidine- 4-carboxamide 566.3358

N2-[4-[4- (isopropylamino)cyclohexen-1- yl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 493.2 359

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]- N2-[4-(piperazin-1-ylmethyl)phenyl]pyrimidine- 2,4-diamine 454.1 360

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [[rac-(1S,6R)-3,9-diazabicyclo[4.2.1]nonan-9- yl]methyl]phenyl]pyrimidine- 2,4-diamine494.2 361

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]- N2-[4-methyl-3-(thiomorpholinomethyl)phenyl] pyrimidine-2,4-diamine 485.1 362

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[3- (piperazin-1-ylmethyl)phenyl]pyrimidine-2,4- diamine 454.2 363

2-amino-N-[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl] ethanesulfonamide 478.1 364

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-thienyl]pyrrolidine-3- carboxamide474.0 365

3-[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazin-1- yl]tetrahydrofuran-2-one 524.2 366

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(morpholinomethyl)phenyl] pyrimidine-2,4-diamine 455.2 367

(2S)-N-methyl-4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxamide 497.1368

azetidin-3-ylmethyl 1-[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxylate 552.1369

methyl 4-[2-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]anilino]ethyl] piperazine-2-carboxylate541.2 370

N2-[4-[(4-isopropylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 496.1 371

4-[[2-[4- (dimethylsulfamoylamino) anilino]pyrimidin-4-yl]amino]-2-(6-methyl- 2-pyridyl)pyrimidine 478.2 372

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(6-oxa-2-azaspiro[3.4]octan-2- ylmethyl)phenyl]pyrimidine- 2,4-diamine481.2 373

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[1-(3-piperidyl)pyrazol-4- yl]pyrimidine-2,4-diamine 429.2 374

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(3-piperidyl)phenyl]pyrimidine- 2,4-diamine 439.2 375

N2-[4-[[(1S,4S)-2,5- diazabicyclo[2.2.1]heptan-2-yl]methyl]phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 466.2 376

N2-[4-[[(2R,5S)-2,5- dimethylpiperazin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 377

N2-[4-(1,4-diazepan-1- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.1 378

N-[[3-(hydroxymethyl)azetidin- 3-yl]methyl]-4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiophene-2- carboxamide 504.0 379

4-methyl-3-[4-[[2-[4-[4- (methylamino)-1- piperidyl]anilino]pyrimidin-4-yl]amino]pyrimidin-2-yl]phenol 483.1 380

N2-[4-[4-(azetidin-3- ylmethyl)piperazin-1-yl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 509.2381

N2-[4-chloro-3-[[rac-(3R,5S)- 3,5-dimethylpiperazin-1-yl]methyl]phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 516.0 382

[rac-(1S)-1-methylpropyl] 4-[[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperazine-2-carboxylate 554.6 383

4-piperidyl 1-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxylate580.2 384

N2-[3-(3,8- diazabicyclo[3.2.1]octan-8- ylmethyl)-4-fluoro-phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 498.1 385

2-(azetidin-3-yl)-N-[3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]acetamide 468.1 386

4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiophene-2- carboxylic acid 406.0 387

N-(azetidin-3-yl)-4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzamide 454.1 388

[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methanol 386.1 389

[5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 3-thienyl]-piperazin-1-yl- methanone474.2 390

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[1-[2-(4-piperidyl)ethyl]pyrazol-4- yl]pyrimidine-2,4-diamine 457.2 391

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- [[rac-(3R,5S)-3,5-dimethylpiperazin-1- yl]methyl]thiazol-2- yl]pyrimidine-2,4-diamine489.1 392

2-[1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] azetidin-3-yl]oxyethanol 485.1 393

N2-[4-[4- [isobutyl(methyl)amino]-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 524.3 394

N2-[4-[4-(dimethylamino)-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 395

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(8-oxa-3-azabicyclo[3.2.1]octan- 3-ylmethyl)phenyl]pyrimidine-2,4-diamine 481.2 396

(4-methylpiperazin-1-yl)-[3-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methanone 482.2 397

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- (4-morpholino-1-piperidyl)phenyl]pyrimidine- 2,4-diamine 524.2 398

N4-[2-(5-chloro-2-fluoro- phenyl)pyrimidin-4-yl]-N2-(4-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 477.1 399

N4-[2-(2- fluorophenyl)pyrimidin-4-yl]- N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 444.2 400

azetidin-3-ylmethyl (3R)-1-[[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 566.1 401

N2-[4-(2-methoxyethyl)phenyl]- N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 414.1 402

azetidin-3-ylmethyl 2-[1-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 567.1 403

[(2R)-4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]methanol 470.1 404

2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- N-[(3R)-pyrrolidin-3-yl]thiazole-4-carboxamide 475.2 405

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] piperidin-4-ol 469.1 406

4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]benzoic acid 399.9 407

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4-morpholinosulfonylphenyl) pyrimidine-2,4-diamine 505.1 408

N4-[2-(5-chloro-2-fluoro- phenyl)pyrimidin-4-yl]- N2-[4-(2-morpholinoethylamino)phenyl] pyrimidine-2,4-diamine 521.1 409

N2-[4-[[(2S,6S)-2,6- dimethylpiperazin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 410

2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 5-morpholino-phenol 457.2 411

methyl 2-[4-[[3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazin-2-yl]acetate526.2 412

N4-[2-(5-chloro-2-fluoro- phenyl)pyrimidin-4-yl]-N2-[4-(4-pyrrolidin-1-yl-1- piperidyl)phenyl]pyrimidine-2,4- diamine 545.1 413

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]pyrrolidine- 3-sulfonamide 504.1 414

[(3R)-1-methylpyrrolidin-3-yl] 1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-4-carboxylate 580.3 415

N4-[2-(5-chloro-2-fluoro- phenyl)pyrimidin-4-yl]-N2-[4-[4-(dimethylamino)-1- piperidyl]phenyl]pyrimidine- 2,4-diamine 520.1 416

N-methyl-N-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine- 4-carboxamide 496.2417

N2-[4-[(1,1-dioxo-1,4-thiazinan- 4-yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 503.1 418

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]- N2-(4-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 440.2 419

N-[7-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2,3-dihydro-1,4-benzodioxin-5-yl]piperidine-4-carboxamide 540.2 420

N-[2-(dimethylamino)ethyl]-3- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzenesulfonamide 506.2 421

N2-(3-amino-4-methyl-phenyl)- N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 385.1 422

(2R)-4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxylic acid484.1 423

3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]benzoic acid 399.9 424

N2-[3-(3,8- diazabicyclo[3.2.1]octan-8- ylmethyl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 480.2 425

N2-[4-(1-isopropylpyrazol-4- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 464.2 426

N4-[2-[6-(difluoromethyl)-5- fluoro-2-pyridyl]pyrimidin- 4-yl]-N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 495.0 427

N-[2-(methylamino)ethyl]-4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzenesulfonamide 492.2 428

(3R)-1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxylicacid 497.2 429

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(4-pyrrolidin-3-ylpiperazin-1- yl)phenyl]pyrimidine-2,4- diamine 509.2430

N-(2-aminoethyl)-3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzenesulfonamide 478.1 431

N2-[4-methoxy-3-[[rac-(3R,5S)- 3,5-dimethylpiperazin-1-yl]methyl]phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 512.2 432

N2-[4-[4- [benzyl(methyl)amino]-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 558.2 433

azetidin-3-ylmethyl 1-[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-3- carboxylate 552.2434

(4-methylpiperazin-1-yl)-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methanone 482.2 435

rac-(3S,5R)-3,5-dimethyl-N-[4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine- 1-carboxamide 511.3436

N4-[2-(5-chloro-2-fluoro- phenyl)pyrimidin-4-yl]-N2-[4-[4-(4-methylpiperazin-1-yl)-1- piperidyl]phenyl]pyrimidine- 2,4-diamine574.2 437

methyl 4-[[2-(4- aminoanilino)pyrimidin-4- yl]amino]-2-(6-methyl-2-pyridyl)pyrimidine-5-carboxylate 429.0 438

N2-[4-[(3-amino-3-methyl- azetidin-1-yl)methyl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 454.2439

5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiophene-2- carboxylic acid 406.3 440

N2-[4-[1-(2-aminoethyl)pyrazol- 4-yl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 465.1 441

4-[[2-[4-[[methyl(4- piperidyl)sulfamoyl]amino] anilino]pyrimidin-4-yl]amino]-2-(6-methyl-2- pyridyl)pyrimidine 547.1 442

methyl 2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]acetate 428.1 443

N-[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperidine- 3-carboxamide 482.1 444

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] piperidine-3-carboxylic acid 497.1 445

N2-(3-chloro-4-piperazin-1-yl- phenyl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 474.1 446

N4-[2-(2- fluorophenyl)pyrimidin-4-yl]- N2-[4-[4-(4-methylpiperazin-1-yl)-1- piperidyl]phenyl]pyrimidine- 2,4-diamine 540.3 447

5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-benzonitrile 465.2 448

4-hydroxy-2-[4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-1- yl]butanoic acid542.2 449

[(1S)-1-methylpropyl] 2-[1-[4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 554.5 450

N2-[4-[4-(benzylamino)-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 544.3 451

methyl 2-[4-[[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazin-2-yl]acetate526.2 452

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-thienyl]piperidine-4- carboxamide488.0 453

N2-[4-[[(2S,5S)-2,5- dimethylpiperazin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 454

1-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperidine-3- carboxylic acid 483.1 455

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-1H-imidazole- 4-sulfonamide 501.0 456

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl]-N-[rac- (3R)-pyrrolidin-3-yl]piperidine-3-carboxamide 565.2 457

N2-[4-(4-cyclopropylpiperazin- 1-yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 480.2 458

N2-[3-(3,8- diazabicyclo[3.2.1]octan-8- ylmethyl)-4-methyl-phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 494.1 459

[(3R)-pyrrolidin-3-yl] 3-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 469.1 460

[(3R)-1-methylpyrrolidin-3-yl] (3R)-1-[[2-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiazol-4-yl]methyl]piperidine- 3-carboxylate 587.2 461

N2-[4-fluoro-3- (thiomorpholinomethyl)phenyl]- N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 489.2 462

methyl 2-[1-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 512.4 463

[(2S)-4-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazin-2-yl]methanol484.1 464

[(3R)-1-methylpyrrolidin-3-yl] 4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]benzoate 483.0 465

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-1-pyrrolidin-3- yl-piperidine-4-carboxamide 551.2 466

[(1S)-1-methylpropyl] 2-[5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetate 554.4467

N2-[4-[[azetidin-3- yl(methyl)amino]methyl]thiazol-2-yl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine461.0 468

methyl 4-[[2-(4- methoxycarbonylanilino) pyrimidin-4-yl]amino]-2-(6-methyl-2-pyridyl) pyrimidine-5-carboxylate 472.0 469

N2-[4-(3,5-dimethylisoxazol-4- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 451.1 470

N2-[4-[[(2R,6S)-2,6- dimethylpiperazin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 471

N2-[4-[4- [(benzylamino)methyl]-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 558.1 472

(1-methylpyrrolidin-3-yl) 1-[4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4-carboxylate 566.2 473

N2-[4-[4-(azetidin-3- yl)piperazin-1-yl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 495.1 474

[1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl]-4- piperidyl]methanol 483.1 475

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 441.4 476

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]- N2-[4-(4-piperidylmethyl)phenyl] pyrimidine-2,4-diamine 453.2 477

1-(azetidin-3-ylmethyl)-3-[3-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]urea 483.1 478

N2-[4-[4-(2- aminoethyl)piperazin-1- yl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 483.3 479

[(3R)-pyrrolidin-3-yl] 1-[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine- 3-carboxylate 552.2480

N2-[4-[(2,2-dimethylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 482.2 481

methyl 1-isopropyl-4-[[4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylate554.2 482

N4-[6-methyl-2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 454.2 483

N2-[4-[4-[2- (methylamino)ethyl]piperazin-1-yl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 497.2 484

1-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methyl] piperidine-4-carboxylic acid 497.1 485

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]- N2-[4-(oxazol-2-ylmethyl)phenyl]pyrimidine- 2,4-diamine 437.1 486

[(3R)-1-methylpyrrolidin-3-yl] (3S)-1-[[4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]methyl]piperidine-3-carboxylate 580.3 487

N2-[4-chloro-3-(3,8- diazabicyclo[3.2.1]octan-8-ylmethyl)phenyl]-N4-[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 514.0 488

(2S)-4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-2- carboxylic acid484.1 489

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- (pyrrolidin-3-ylmethyl)phenyl]pyrimidine- 2,4-diamine 439.2 490

3-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]anilino]methyl] azetidin-3-ol 456.2 491

methyl 2-[4-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazin-2- yl]acetate 512.2 492

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(1,2,3,4-tetrahydroquinolin-6- yl)pyrimidine-2,4-diamine 411.2 493

N2-[4-[(3-methoxy-3-methyl- azetidin-1-yl)methyl]phenyl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 469.1494

N2-[4-(4-isobutylpiperazin-1- yl)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 496.2 495

N-(azetidin-3-ylmethyl)-1-[[4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxamide565.3 496

N2-[4-[azetidin-3- yl(methyl)amino]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 440.1 497

N-methyl-N-[3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 496.2498

N-(2-hydroxyethyl)-2-[2-(6- methyl-2-pyridyl)-6-[[2-(4-piperazin-1-ylanilino)pyrimidin- 4-yl]amino]pyrimidin-4- yl]acetamide541.1 499

[(3R)-1-methylpyrrolidin-3-yl] 2-[5-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]-2-piperazin-1-yl-phenyl]acetate 581.1 500

N2-(5-amino-3-thienyl)-N4-[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 377.1 501

N2-[4-[[3- (dimethylamino)azetidin-1- yl]methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.3 502

N2-(2-fluoro-4-morpholino- phenyl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 459.1 503

N2-[4- (methylaminomethyl)phenyl]- N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 399.2 504

[(3R)-1-methylpyrrolidin-3-yl] 4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 489.0 505

[3-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]-piperazin-1-yl- methanone 468.1 506

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[rac-(2R)-2-methylpiperazin-1- yl]phenyl]pyrimidine-2,4- diamine 454.1507

[(3S)-1-methylpyrrolidin-3-yl] 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 489.1 508

[(3S)-1-methylpyrrolidin-3-yl] (3R)-1-[[2-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiazol-4-yl]methyl]piperidine-3- carboxylate 587.1 509

4-methyl-N-[3-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-1- carboxamide 497.1510

[2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazol-4-yl]- piperazin-1-yl-methanone 475.0 511

N-[2-methoxy-4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 512.2512

N2-(1H-benzotriazol-5-yl)-N4- [2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 397.1 513

2-[[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]methoxy] ethanol 430.2 514

N2-(4-amino-3-methyl-phenyl)- N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 385.1 515

N2-(3-methyl-4-piperazin-1-yl- phenyl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 454.1 516

methyl 1-[[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperidine-3-carboxylate511.1 517

methyl 3-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]propanoate 442.1 518

azetidin-3-ylmethyl 1-[[4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] pyrrolidine-3-carboxylate552.1 519

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[3- [[rac-(3R,5S)-3,5-dimethylpiperazin-1- yl]methyl]phenyl]pyrimidine- 2,4-diamine 482.1 520

N-[3-(hydroxymethyl)pyrrolidin- 3-yl]-2-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiazole-4-carboxamide 505.1 521

N2-[3-[(4-methylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 468.2 522

[(3R)-1-methylpyrrolidin-3-yl] 5-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-3-carboxylate 489.2 523

isobutyl 4-[[4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]methyl] piperazine-2-carboxylate554.2 524

N2-(2-aminoindan-5-yl)-N4-[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]pyrimidine-2,4-diamine 411.1 525

N2-[4-[4- [(isobutylamino)methyl]-1- piperidyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 524.1 526

N2-(2,3-dihydro-1,4- benzodioxin-6-yl)-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 414.1 527

N2-[4-(4-cyclopropylpiperazin- 1-yl)sulfonylphenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 544.0 528

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[rac-(3S)-3-methylpiperazin-1- yl]phenyl]pyrimidine-2,4- diamine 454.1529

ethyl 2-[2-(6-methyl-2-pyridyl)- 6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4-yl]acetate 526.1 530

azetidin-3-yl 2-[5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-piperazin-1-yl-phenyl]acetate 553.1531

4-isopropyl-N-[3-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperazine-1- carboxamide 525.2532

5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiophene-3-carboxylic acid 406.2 533

N2-[4-[(4-isobutylpiperazin-1- yl)methyl]phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 510.3 534

2-[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl]piperazin- 1-yl]ethanol 484.3 535

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4- (1H-pyrazol-4-yl)phenyl]pyrimidine-2,4- diamine 422.0 536

[(3R)-3- (hydroxymethyl)piperazin-1-yl]- [4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]- 2-thienyl]methanone504.1 537

N2-[4-(azetidin-3- ylmethylamino)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 440.2 538

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[1-(oxetan-3-yl)pyrazol-4- yl]phenyl]pyrimidine- 2,4-diamine 478.1 539

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4-piperazin-2-ylphenyl)pyrimidine- 2,4-diamine 440.1 540

N4-[2-(5-chloro-2,4-difluoro- phenyl)-5-cyclopropyl-pyrimidin-4-yl]-N2-(4-piperazin- 1-ylphenyl)pyrimidine-2,4- diamine535.1 541

2-[4-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]phenyl] sulfonylpiperazin- 1-yl]ethanol 458.0 542

N4-[2-(5-fluoro-6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-[4-(4-methylpiperazin-1-yl)-1- piperidyl]phenyl]pyrimidine-2,4- diamine555.3 543

[(3R)-pyrrolidin-3-yl] 4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 475.2 544

[(3S)-pyrrolidin-3-yl] 4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 475.1 545

azetidin-3-ylmethyl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 470.1 546

[(3R)-pyrrolidin-3-yl] 5-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 470.1 547

[(3S)-pyrrolidin-3-yl] 5-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 470.1 548

azetidin-3-ylmethyl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 470.1 549

[(3R)-pyrrolidin-3-yl] 5-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 470.1 550

[(3S)-pyrrolidin-3-yl] 5-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 470.2 551

2-piperazin-1-ylethyl 2-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4-carboxylate 519.2 552

2-piperazin-1-ylethyl 5-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 513.2 553

2-piperazin-1-ylethyl 3-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]benzoate 512.2 554

2-piperazin-1-ylethyl 4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 518.1 555

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(5- morpholinothiazol-2-yl)pyrimidine-2,4-diamine 448.2 556

N4-[2-(5-chloro-2,4-difluoro- phenyl)pyrimidin-4-yl]-N2-(4-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 495.1 557

3-azabicyclo[3.1.0]hexan-1- ylmethyl 5-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-2-carboxylate 496.2 558

2-(4-piperidyl)ethyl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 512.3 559

6-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-3- carboxylic acid 401.1 560

2-(azetidin-3-ylamino)ethyl 4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 504.1 561

4-piperidyl 5-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 484.2 562

[(1R)-1-(4-piperidyl)ethyl] 5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 512.2 563

(4-aminocyclohexyl)methyl 4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 517.1 564

azetidin-3-ylmethyl 6-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 470.2 565

4-piperidyl 1-[[5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 3-pyridyl]methyl]azetidine-3-carboxylate 553.2 566

(4-amino-4-methyl-cyclohexyl) 4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 517.1 567

[(3R,6R)-6-methyl-3-piperidyl] 5-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-2-carboxylate 498.2 568

2-[(3R)-3-aminopyrrolidin-1- yl]ethyl 4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 518.1 569

1-[[5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 2-pyridyl]methyl]azetidine-3- carboxylicacid 470.1 570

[(3R)-pyrrolidin-3-yl]methyl 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 484.2 571

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(1,2,3,6-tetrahydropyridin-4- yl)thiazol-2-yl]pyrimidine- 2,4-diamine444.2 572

(4-amino-4-methyl-cyclohexyl) 4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 517.2 573

[(3S)-pyrrolidin-3-yl] 1-[[5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 3-pyridyl]methyl]azetidine-3-carboxylate 539.2 574

[(3R)-pyrrolidin-3-yl] 6-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 470.2 575

[(3S)-pyrrolidin-3-yl] 6-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine- 3-carboxylate 470.1 576

(1-acetylazetidin-3-yl)methyl 4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 517.1 577

[(2R)-pyrrolidin-2-yl]methyl 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 484.1 578

[(2R)-2-piperidyl]methyl 2-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-5- carboxylate 504.1 579

azetidin-3-yl 2-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-4- carboxylate 456.1 580

[(3R)-3-piperidyl]methyl 2-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4- carboxylate 504.2 581

2-piperazin-1-ylethyl 6-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 513.2 582

[(3S)-pyrrolidin-3-yl]methyl 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 484.3 583

[(2R)-pyrrolidin-2-yl]methyl 4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 489.1 584

[(3R)-pyrrolidin-3-yl]methyl 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-2-carboxylate 484.1 585

4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazole-2- carboxylic acid 407.1 586

[(2R)-2-piperidyl]methyl 2-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4- carboxylate 504.3 587

4-piperidylmethyl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2- carboxylate 498.1 588

[(2R)-2-piperidyl]methyl 4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 503.1 589

2-[(3R,5S)-3,5- dimethylpiperazin-1-yl]ethyl 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 541.2 590

(4-aminocyclohexyl)methyl 4- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 517.2 591

4-piperidyl 4-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 488.9 592

[(2R)-pyrrolidin-2-yl]methyl 2- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazole-5-carboxylate 490.1 593

[(2R)-pyrrolidin-2-yl]methyl 2- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazole-4-carboxylate 490.1 594

[(3R)-3-piperidyl] 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2- carboxylate 484.1 595

(3R)-3-piperidyl]methyl 5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 498.1 596

azepan-4-yl 5-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2- carboxylate 498.2 597

[(2S)-2-piperidyl]methyl 5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2- carboxylate 498.1 598

2-[(3R)-3-amino-1- piperidyl]ethyl 4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 532.1 599

2-(3,4,6,7,8,8a-hexahydro-1H- pyrrolo[1,2-a]pyrazin-2-yl)ethyl4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiophene- 2-carboxylate 558.2 600

[(3S)-3-piperidyl]methyl 4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 503.1 601

[(3R)-pyrrolidin-3-yl] 2-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-4- carboxylate 470.1 602

2-(azetidin-3-yl)ethyl 4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 489.1 603

1-[[5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 3-pyridyl]methyl]azetidine-3- carboxylicacid 470.1 604

N2-[5-[(3R,5S)-3,5- dimethylpiperazin-1-yl]thiazol-2-yl]-N4-[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine475.1 605

[(2R)-morpholin-2-yl]methyl 5- [[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 500.1 606

4-piperidylmethyl 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 498.2 607

4-piperidylmethyl 4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 503.1 608

azetidin-3-ylmethyl 2-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-4-carboxylate 470.1 609

N2-[5-(2,5-dihydro-1H-pyrrol-3- yl)thiazol-2-yl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 430.2 610

4-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-2-carboxylic acid 401.2 611

[(3R)-pyrrolidin-3-yl]methyl 2- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazole-4-carboxylate 490.1 612

[(2S)-pyrrolidin-2-yl]methyl 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-2-carboxylate 484.1 613

2-(3,4,6,7,8,8a-hexahydro-1H- pyrrolo[1,2-a]pyrazin-2-yl)ethyl5-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-2-carboxylate 553.0 614

azetidin-3-ylmethyl 1-[[5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 3-pyridyl]methyl]azetidine-3-carboxylate 539.2 615

4-piperidylmethyl 2-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4-carboxylate 504.3 616

azetidin-3-ylmethyl 4-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 470.1 617

[(3S)-3-piperidyl] 5-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 484.4 618

[(2R)-pyrrolidin-2-yl]methyl 5- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-3-carboxylate 484.1 619

2-[(3S)-3-amino-1- piperidyl]ethyl 4-[[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 532.1 620

[(3S)-3-piperidyl]methyl 5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 498.2 621

[(1R,3S)-3- aminocyclohexyl]methyl 4-[[4- [[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 517.1 622

azetidin-3-yl 6-[[4-[[2-(6- methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 456.1 623

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[5-(1,2,3,6-tetrahydropyridin-4- yl)thiazol-2-yl]pyrimidine-2,4- diamine444.3 624

2-piperazin-1-ylethyl 2-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-4-carboxylate 513.2 625

[(3R)-3-piperidyl]methyl 5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 498.1 626

N2-[4-(2,5-dihydro-1H-pyrrol-3- yl)thiazol-2-yl]-N4-[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]pyrimidine-2,4-diamine 430.2 627

2-[(3S,5R)-3,5- dimethylpiperazin-1-yl]ethyl 4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 546.1 628

[(2R)-2-piperidyl]methyl 5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 498.2 629

1-[[2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazol-4- yl]methyl]azetidine-3- carboxylic acid 476.1 630

[(3R)-pyrrolidin-3-yl]methyl 2- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]thiazole-5-carboxylate 490.1 631

(3-aminocyclobutyl) 4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 475.1 632

[(3S)-pyrrolidin-3-yl] 2-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-4-carboxylate 470.1 633

4-piperidyl 2-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-4-carboxylate 484.1 634

4-piperidyl 6-[[4-[[2-(6-methyl- 2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3-carboxylate 484.2 635

[(1R,3R)-3- aminocyclohexyl]methyl 4-[[4- [[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 517.1 636

2-[[4-[[2-(6-methyl-2- pyridyl)pyrimidin-4- yl]amino]pyrimidin-2-yl]amino]pyridine-4- carboxylic acid 401.2 637

[(2S)-pyrrolidin-2-yl]methyl 4- [[4-[[2-(6-methyl-2-pyridyl)pyrimidin-4- yl]amino]pyrimidin-2- yl]amino]thiophene-2-carboxylate 489.1 638

(3-aminocyclobutyl) 4-[[4-[[2- (6-methyl-2-pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 475.1 639

[(2S)-2-piperidyl]methyl 4-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylate 503.1 640

N4-[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(5-piperazin-1-ylthiazol-2- yl)pyrimidine-2,4-diamine 447.2 641

[(2R)-2-piperidyl]methyl 5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3- carboxylate 498.2 642

[(3R)-pyrrolidin-3-yl] 1-[[5-[[4- [[2-(6-methyl-2- pyridyl)pyrimidin-4-yl]amino]pyrimidin-2-yl]amino]- 3-pyridyl]methyl]azetidine-3-carboxylate 539.2

In some embodiments, exemplary compounds may include, but are notlimited to, a compound or salt thereof selected from Table 2.

TABLE 2 No. Structure Chemical Name [M + H]⁺ 1000

N4-[6-(6-methyl-2-pyridyl)-2- pyridyl]-N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 440.1 1001

N4-[6-(6-methyl-2-pyridyl)-1H- pyrrolo[2,3-b]pyridin-4-yl]-N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 479.3 1002

N4-[5-(6-methyl-2-pyridyl)-3H- triazolo[4,5-d]pyrimidin-7-yl]-N2-(4-morpholinophenyl)pyrimidine- 2,4-diamine 482.1 1003

2-(6-methyl-2-pyridyl)-6-[[2-(4- morpholinoanilino)pyrimidin-4-yl]amino]pyridine-3- carbonitrile 465.2 1004

ethyl 2-[2-(6-methyl-2-pyridyl)-6- [[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4-yl]acetate 526.1 1005

N4-[6-methyl-2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-(4-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 454.2 1006

N-ethyl-2-[2-(6-methyl-2-pyridyl)- 6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4-yl] acetamide 525.1 1007

N-(2-hydroxyethyl)-2-[2-(6-methyl- 2-pyridyl)-6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4-yl] acetamide 541.1 1008

N-methyl-2-[2-(6-methyl-2-pyridyl)- 6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4-yl] acetamide 511.2 1009

N-isobutyl-2-[2-(6-methyl-2- pyridyl)-6-[[2-(4-piperazin-1-ylanilino)pyrimidin-4- yl]amino]pyrimidin-4-yl] acetamide 553.2 1010

methyl 6-[[2-[4-(4-methylpiperazin- 1-yl)anilino]pyrimidin-4-yl]amino]-2-(6-methyl-2-pyridyl)pyrimidine- 4-carboxylate 512.2 1011

N4-[6-methyl-2-(6-methyl-2- pyridyl)pyrimidin-4-yl]-N2-[4-(4-methylpiperazin-1- yl)phenyl]pyrimidine-2,4-diamine 468.1 1012

N4-[2-(6-methyl-2-pyridyl)-6-(1H- triazol-4-yl)pyrimidin-4-yl]-N2-(4-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 507.2 1013

N4-[2-(5-chloro-2-fluoro-phenyl)-5- cyclopropyl-pyrimidin-4-yl]-N2-(4-piperazin-1-ylphenyl)pyrimidine- 2,4-diamine 517.1 1014

N4-[2-(5-chloro-2,4-difluoro- phenyl)-5-cyclopropyl-pyrimidin-4-yl]-N2-(4-piperazin-1- ylphenyl)pyrimidine-2,4-diamine 535.1 1015

N2-(4-aminophenyl)-N4-[2-(6- methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]pyrimidine- 2,4-diamine 410.1 1016

N2-[4-(azetidin-3- ylmethylamino)phenyl]-N4-[2-(6-methyl-2-pyridyl)pyrrolo[2,1- f][1,2,4]triazin-4-yl]pyrimidine-2,4-diamine 479.1 1017

3-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]benzoic acid 439.1 1018

2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]acetic acid 453.2 1019

methyl 2-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]acetate 467.1 1020

6-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f]]1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-3- carboxylic acid 440.0 1021

4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]benzoic acid 439.1 1022

2-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]thiazole-4- carboxylic acid 446.0 1023

4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-2- carboxylic acid 445.0 1024

methyl 5-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]pyridine-2-carboxylate 454.0 1025

5-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]thiophene-3- carboxylic acid 444.9 1026

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]azetidine-3- carboxamide 493.11027

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]pyrrolidine-3- carboxamide 507.11028

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-4- carboxamide 521.21029

2-(azetidin-3-yl)-N-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]acetamide507.1 1030

3-amino-N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl] cyclobutanecarboxamide 507.11031

3-(hydroxymethyl)-N-[4-[[4-[[2-(6- methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4- yl]amino]pyrimidin-2- yl]amino]phenyl]azetidine-3-carboxamide 523.1 1032

N-[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]piperidine-3- carboxamide 521.21033

2,7-diazaspiro[3.5]nonan-7-yl-[4- [[4-[[2-(6-methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin- 4-yl]amino]pyrimidin-2-yl]amino]phenyl]methanone 547.1 1034

N-(azetidin-3-yl)-4-[[4-[[2-(6- methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4- yl]amino]pyrimidin-2- yl]amino]benzamide 493.0 1035

N-(azetidin-3-ylmethyl)-4-[[4-[[2- (6-methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4- yl]amino]pyrimidin-2- yl]amino]benzamide 507.1 1036

4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2-yl]amino]- N-pyrrolidin-3-yl-benzamide 507.1 1037

4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2-yl]amino]- N-(4-piperidyl)benzamide 521.1 1038

[4-[[4-[[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]amino]pyrimidin-2- yl]amino]phenyl]-piperazin-1-yl- methanone 507.01039

azetidin-3-ylmethyl 4-[[4-[[2-(6- methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4- yl]amino]pyrimidin-2- yl]amino]benzoate 508.1 1040

N2-[4-(azetidin-3-ylamino)phenyl]- N4-[2-(6-methyl-2-pyridyl)pyrrolo[2,1-f][1,2,4]triazin- 4-yl]pyrimidine-2,4-diamine 465.11041

N4-[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin-4-yl]-N2-[4-(pyrrolidin-3- ylamino)phenyl]pyrimidine- 2,4-diamine 479.11042

N4-[2-(6-methyl-2- pyridyl)pyrrolo[2,1-f][1,2,4]triazin- 4-yl]-N2-[4-(4-piperidylamino)phenyl] pyrimidine-2,4-diamine 493.1 1043

methyl 4-[[2-(4- methoxycarbonylanilino)pyrimidin-4-yl]amino]-2-(6-methyl-2- pyridyl)pyrimidine-5-carboxylate 472.0 1044

4-[[2-(4-aminoanilino)pyrimidin-4- yl]amino]-2-(6-methyl-2-pyridyl)pyrimidine-5- carboxylic acid 415.0 1045

methyl 4-[[2-(4- aminoanilino)pyrimidin-4- yl]amino]-2-(6-methyl-2-pyridyl)pyrimidine-5-carboxylate 429.0

Methods

In some aspects, the present disclosure provides a method of inhibitingTGFβ signaling, comprising contacting a cell with an effective amount ofa compound disclosed herein, such as a compound of Formula (I), (I′),(I-A), (I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or(I′-E). In some embodiments, the present disclosure provides a method ofinhibiting ALK5, comprising contacting ALK5 with an effective amount ofa compound disclosed herein. Inhibition of ALK5 or TGFβ signaling can beassessed by a variety of methods known in the art. Non-limiting examplesinclude a showing of (a) a decrease in kinase activity of ALK5; (b) adecrease in binding affinity between the TGFβ/TGFβ-RII complex and ALK5;(c) a decrease in the levels of phosphorylated intracellular signalingmolecules downstream in the TGFβ signaling pathway, such as a decreasein pSMAD2 or pSMAD3 levels; (d) a decrease in binding of ALK5 todownstream signaling molecules, such as SMAD2 and SMAD3; and/or (e) anincrease in ATP levels or a decrease in ADP levels. Kits andcommercially available assays can be utilized for determining one ormore of the above.

In some aspects, the present disclosure provides a method of treating anALK5-mediated disease or condition in a subject, comprisingadministering to the subject a therapeutically effective amount of acompound disclosed herein. In some embodiments, the disease or conditionis selected from fibrosis and cancer. In some embodiments, the diseaseor condition is pulmonary fibrosis, such as idiopathic pulmonaryfibrosis or virus-induced fibrosis. In some embodiments, the disease orcondition is intestinal fibrosis. In some embodiments, the disease orcondition is alopecia. In some embodiments, the disease is aneurodegenerative disease, such as Alzheimer's disease. In someembodiments, the present disclosure provides a method of reversingsymptoms of aging. For example, the method may enhance neurogenesis,reduce neuroinflammation, improve cognitive performance, regenerateliver tissue, and reduce p16 levels.

In some aspects, the present disclosure provides a method of treatingfibrosis, comprising administering to a patient an effective amount of acompound disclosed herein. In some embodiments, the fibrosis is mediatedby ALK5. In some embodiments, the fibrosis is selected from systemicsclerosis, systemic fibrosis, organ-specific fibrosis, kidney fibrosis,pulmonary fibrosis, liver fibrosis, portal vein fibrosis, skin fibrosis,bladder fibrosis, intestinal fibrosis, peritoneal fibrosis,myelofibrosis, oral submucous fibrosis, and retinal fibrosis. In someembodiments, the fibrosis is pulmonary fibrosis, such as idiopathicpulmonary fibrosis (IPF), familial pulmonary fibrosis (FPF),interstitial lung fibrosis, fibrosis associated with asthma, fibrosisassociated with chronic obstructive pulmonary disease (COPD),silica-induced fibrosis, asbestos-induced fibrosis orchemotherapy-induced lung fibrosis. In some embodiments, the fibrosis isidiopathic pulmonary fibrosis (IPF). In some embodiments, the fibrosisis TGF-β-mediated pulmonary fibrosis. In some embodiments, the patienthas been diagnosed with acute respiratory distress syndrome (ARDS). Insome embodiments, the fibrosis is acute fibrosis. In some embodiments,the fibrosis is chronic fibrosis.

In some aspects, the present disclosure provides a method of treatingpulmonary fibrosis induced by a viral infection, comprisingadministering to a patient an effective amount of a compound disclosedherein. The pulmonary fibrosis may be induced by an erythrovirus, adependovirus, a papillomavirus, a polyomavirus, a mastadenovirus, analphaherpesvirinae, a varicellovirus, a gammaherpesvirinae, abetaherpesvirinae, a roseolovirus, an orthopoxvirus, a parapoxvirus, amolluscipoxvirus, an orthohepadnavirus, an enterovirus, a rhinovirus, ahepatovirus, an aphthovirus, a calicivirus, an astrovirus, analphavirus, a rubivirus, a flavivirus, a Hepatitis C virus, a reovirus,an orbivirus, a rotavirus, an influenzavirus A, an influenzavirus B, aninfluenzavirus C, a paramyxovirus, a morbillivirus, a rubulavirus, apneumovirus, a vesiculovirus, a lyssavirus, a bunyavirus, a hantavirus,a nairovirus, a phlebovirus, a coronavirus, an arenavirus, aBLV-HTLV-retrovirus, a lentivirinae, a spumavirinae or a filovirus. Insome embodiments, the fibrosis is virus-induced fibrosis, such asvirus-induced pulmonary fibrosis. In some embodiments, the fibrosis isselected from EBV-induced pulmonary fibrosis, CMV-induced pulmonaryfibrosis, herpesvirus-induced pulmonary fibrosis and coronavirus-inducedpulmonary fibrosis. In some embodiments, the fibrosis is selected fromEBV-induced pulmonary fibrosis, CMV-induced pulmonary fibrosis,HHV-6-induced pulmonary fibrosis, HHV-7-induced pulmonary fibrosis,HHV-8-induced pulmonary fibrosis, H5N1 virus-induced pulmonary fibrosis,SARS-CoV-induced pulmonary fibrosis, MERS-CoV-induced pulmonary fibrosisand SARS-CoV-2-induced pulmonary fibrosis. In some embodiments, thepulmonary fibrosis is coronavirus-induced pulmonary fibrosis. In someembodiments, the pulmonary fibrosis is SARS-CoV-2-induced pulmonaryfibrosis. In some embodiments, the pulmonary fibrosis isCOVID-19-induced pulmonary fibrosis.

In some aspects, the present disclosure provides a method of treatingacute lung injury (ALI), comprising administering to a patient aneffective amount of a compound disclosed herein. In some embodiments,the present disclosure provides a method of treating acute respiratorydistress syndrome (ARDS), comprising administering to a patient aneffective amount of a compound disclosed herein. The ARDS may be in theearly acute injury phase or the fibroproliferative phase. In someembodiments, the ARDS is fibroproliferative ARDS. In some embodiments,the present disclosure provides a method of treating fibrosis resultingfrom ARDS, comprising administering to a patient an effective amount ofa compound disclosed herein. The fibrosis resulting from ARDS may bepulmonary fibrosis. In some embodiments, the present disclosure providesa method of treating fibrosis resulting from ALI, comprisingadministering to a patient an effective amount of a compound disclosedherein. The fibrosis resulting from ALI may be pulmonary fibrosis.

In some aspects, the present disclosure provides a method of treatingintestinal fibrosis, comprising administering to a patient an effectiveamount of a compound disclosed herein. In some embodiments, theintestinal fibrosis is mediated by ALK5. In some embodiments, thecompound is administered in an amount effective to delay progression of,reduce the incidence of, or reduce the degree of one or morecharacteristics associated with intestinal fibrosis. In someembodiments, the compound is administered, either in a single dose orover multiple doses, in an amount effective to reverse establishedfibrosis.

In some aspects, the present disclosure provides a method of treatingcancer, comprising administering to a patient an effective amount of acompound disclosed herein. In some embodiments, the cancer is mediatedby ALK5. In some embodiments, the cancer is selected from breast cancer,colon cancer, prostate cancer, lung cancer, hepatocellular carcinoma,glioblastoma, melanoma and pancreatic cancer. In some embodiments, thecancer is lung cancer, such as non-small cell lung cancer. In someaspects, the present disclosure provides a method of treating cancer,such as non-small cell lung cancer, comprising administering to apatient an effective amount of a compound disclosed herein and animmunotherapeutic agent. In some embodiments, the cancer is stage IIInon-small cell lung cancer. In some embodiments, the method furthercomprises administering radiation to the patient. In some embodiments,the immunotherapeutic agent is a PD-1 inhibitor or a CTLA-4 inhibitor.In some embodiments, the immunotherapeutic agent is selected fromatezolizumab, avelumab, nivolumab, pembrolizumab, durvalumab, BGB-A317,tremelimumab and ipilimumab. In some embodiments, the immunotherapeuticagent is selected from pembrolizumab and durvalumab.

The compounds described herein, including compounds of Formula (I),(I′), (I-A), (I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E)or (I′-E), are ALK5 inhibitors that limit the activity of TGFβ. TGFβ isone of several factors involved in the initiation and development offibrotic diseases throughout the body. As such, the compounds of thedisclosure are expected to be useful for the treatment, preventionand/or reduction of fibrosis in a patient by administering atherapeutically effective amount of a compound disclosed herein. Byinhibiting ALK5, the compound is expected to potentiate the formation offibrosis in areas of the body that suffer from excessive deposition ofthe extracellular matrix. Those areas are described below.

Systemic Fibrotic Diseases

Systemic sclerosis (SSc) is an autoimmune disorder that affects the skinand internal organs and results in autoantibody production, vascularendothelial activation of small blood vessels, and tissue fibrosis as aresult of fibroblast dysfunction. Transforming growth factor β (TGF-β)has been identified as a regulator of pathological fibrogenesis in SSc(Ayers, N. B., et al., Journal of Biomedical Research, 2018, 32(1), pp.3-12). According to the authors, “understanding the essential role TGF-βpathways play in the pathology of systemic sclerosis could provide apotential outlet for treatment and a better understanding of this severedisease.” In some embodiments, the present disclosure provides a methodof treating SSc, comprising administering to a subject an effectiveamount of a compound disclosed herein.

Multifocal fibrosclerosis (MF) and idiopathic multifocal fibrosclerosis(IMF) are disorders characterized by fibrous lesions at varying sitesand include retroperitoneal fibrosis, mediastinal fibrosis and Riedel'sthyroiditis. Both multifocal fibrosclerosis and idiopathic multifocalfibrosclerosis are considered to be an outcome of IgG₄-associatedfibrosis/disease and TGF-β is believed to be one factor involved in theinitiation and development of fibrosis (Pardali, E., et. al., Int. J.Mol. Sci., 18, 2157, pp. 1-22). In some embodiments, the presentdisclosure provides a method of treating multifocal fibrosclerosis oridiopathic multifocal fibrosclerosis, comprising administering to asubject an effective amount of a compound disclosed herein.

In some embodiments, the present disclosure provides a method oftreating nephrogenic systemic fibrosis, comprising administering to asubject an effective amount of a compound disclosed herein. Nephrogenicsystemic fibrosis is a rare disease occurring mainly in people withadvanced kidney failure with or without dialysis. In a study performedby Kelly et al. (J. Am. Acad. Dermatol., 2008, 58, 6, pp. 1025-1030), itwas shown that TGF-β, as well as Smad 2/3, appear to be associated withfibrosis seen in nephrogenic systemic fibrosis.

Sclerodermatous graft-versus-host disease (GVHD) is a prevalentcomplication of allogeneic hematopoietic stem cell graft appearing twoto three months after allogeneic bone marrow transplantation. Thedisease results in production of autoantibodies and fibrosis of skin andinner organs. Using a murine cutaneous GVHD model, it has been shownthat progression of early skin and lung disease can be inhibited withTGF-β neutralizing antibodies (McCormick, L. L., et al., J. Immunol.,1999, 163, pp. 5693-5699). In some embodiments, the present disclosureprovides a method of treating sclerodermatous GVHD, comprisingadministering to a subject an effective amount of a compound disclosedherein.

Organ-Specific Fibrotic Diseases

Cardiac fibrosis refers to the abnormal thickening of heart valves dueto the abnormal proliferation of cardiac fibroblasts resulting in excessdeposition of ECM in heart muscle. Fibroblasts secrete collagen, whichserves as structural support for the heart. However, when collagen isexcessively secreted in the heart, wall and valve thickening can resultin tissue build-up on the tricuspid and pulmonary valves. This in turncauses loss of flexibility and ultimately valvular dysfunction leadingto heart failure. A specific type of cardiac fibrosis ishypertension-associated cardiac fibrosis as described by J. Diez (J.Clin. Hypertens., 2007, July 9(7), pp. 546-550). According to Diez,changes in the composition of cardiac tissue develop in hypertensivepatients with left ventricular hypertrophy and lead to structuralremodeling of the heart tissue. One change relates to the disruption ofthe equilibrium between the synthesis and degradation of collagen typesI and III molecules, resulting in excessive accumulation of collagenfibers in the heart tissue. Other types of cardiac fibrosis includepost-myocardial infarction and Chagas disease-induced myocardialfibrosis. In Chagas disease, transforming growth factor β1 (TGF-β1) hasbeen implicated in Chagas disease physiopathology, where animal modelssuggest that the TGF-β1-pathway is up-regulated during infection(Araujo-Jorge, T. C., et al., Clin. Pharmacol. Ther., 2012, 92(5), pp.613-621; Curvo, E., Mem Inst Oswaldo Cruz, 2018, Vol. 113(4), e170440,pp. 1-8). In some embodiments, the present disclosure provides a methodof treating various forms of cardiac fibrosis, such ashypertension-associated cardiac fibrosis, post-myocardial infarction orChagas disease-induced myocardial fibrosis, comprising administering toa subject an effective amount of a compound disclosed herein.

Renal fibrosis encompasses a variety of disorders associated with theaberrant expression and activity of TGF-β, including, but not limitedto, diabetic and hypertensive nephropathy, urinary tractobstruction-induced kidney fibrosis, inflammatory/autoimmune-inducedkidney fibrosis, aristolochic acid nephropathy, progressive kidneyfibrosis, and polysystic kidney disease. As discussed above, fibrosisinvolves an excess accumulation of the ECM, which in turn causes loss offunction when normal tissue is replaced with scar tissue (Wynn, T. A., JClin Invest., 2007, 117, pp. 524-529). As early as 2005, ALK5 inhibitorswere being studied in models for renal disease (Laping, N.J., CurrentOpinion in Pharmacology, 2003, 3, pp. 204-208). In some embodiments, thepresent disclosure provides a method of treating renal fibrosis,comprising administering to a subject an effective amount of a compounddisclosed herein.

One fibrotic disease that has been particularly difficult to treat isidiopathic pulmonary fibrosis (IPF). IPF is a chronic, progressive andfatal fibrotic lung disease with survival only improved by lungtransplantation. Current oral therapies such as nintedanib andpirfenidone have been shown to slow the progression of the disease, buthave adverse effects that lead to discontinuation and lack of complianceby the patient. Although there are other therapies in developmenttargeting various pathways, an unmet need remains for patients with IPF.

Although ALK5 is an important and known component in the fibroticdisease pathway, the efficacy of ALK5 inhibitors in IPF have not beenrealized due to systemic adverse effects, especially in the heart. Thus,one of the goals of this disclosure is to develop ALK5 inhibitors withhigh lung selectivity and rapid clearance. One preferred embodiment ofthis disclosure is to treat patients with idiopathic pulmonary fibrosiswith a compound described herein, for example, by once or twice dailyadministration of inhalable ALK5 inhibitor having minimal systemicexposure. The inhaled ALK5 inhibitor may be administered as amonotherapy or co-dosed with other orally available IPF therapies. Insome embodiments, the present disclosure provides a method of treatingidiopathic pulmonary fibrosis, comprising administering to a subject aneffective amount of a compound disclosed herein. In some embodiments,the compound is administered by inhalation.

Familial pulmonary fibrosis is a hereditary disease where two or morefamily members have confirmed IPF. In some embodiments, the presentdisclosure provides a method of treating familial pulmonary fibrosis,comprising administering to a subject an effective amount of a compounddisclosed herein.

Pulmonary fibrosis is a typical clinical feature associated with viralinfection, such as SARS and COVID-19. SARS-mediated TGF-β signaling hasbeen shown to promote fibrosis and block apoptosis of SARS-CoV-infectedhost cells (Zhao, X. et al., J. Biol. Chem., 2008, 283(6), pp.3272-3280). Increased TGF-β expression was similarly observed inpatients infected with SARS-CoV-2, ultimately leading to the developmentof pulmonary fibrosis. TGF-β signaling mediated by SARS-CoV-2 canpromote fibroblast proliferation and myofibroblast differentiation andblock host cell apoptosis. (Xiong, Y. et al., Emerging Microbes &Infections, 2020, 9(1), pp. 761-770). Compounds of the presentdisclosure are expected to inhibit increased TGF-β signaling mediated byviral infection and prevent, halt, slow or reverse the progression ofpulmonary fibrosis associated with the infection. Accordingly, in someembodiments, the present disclosure provides a method of treatingpulmonary fibrosis induced by a viral infection, comprisingadministering to a subject an effective amount of a compound disclosedherein. In some embodiments, the pulmonary fibrosis is induced bySARS-CoV or SARS-CoV-2. In some embodiments, the compound isadministered by inhalation.

Chronic lung disease, such as interstitial lung disease (ILD), chronicobstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis(IPF), may lead to pulmonary hypertension (PH). Pulmonary hypertensionis a progressive disease characterized by high blood pressure in thelungs. The World Health Organization (WHO) has defined fiveclassifications of PH (WHO Group I: Pulmonary arterial hypertension(PAH); WHO Group II: Pulmonary hypertension due to left heart disease;WHO Group III: Pulmonary hypertension due to lung disease and/orhypoxia; WHO Group IV: Chronic thromboembolic pulmonary hypertension(CTEPH); and WHO Group V: Pulmonary hypertension with unclearmultifactorial mechanisms). TGF-β signaling has been implicated in thepathogenesis of PH. Moreover, inhibition of ALK5 in a monocrotaline(MCT) model of severe PH was shown to attenuate the development of PHand reduce pulmonary vascular remodeling in a dose-dependent manner,namely by reducing RV systolic pressure, reducing RV diastolic pressure,increasing cardiac output and reducing RV hypertrophy (Zaiman, A. L.; etal., Am. J. Respir. Crit. Care Med., 2008, 177, pp. 896-905). Compoundsof the present disclosure are expected to inhibit TGF-β signaling inlung tissue and prevent, halt, slow or reverse the progression of PH,particularly in WHO Group III PH. Accordingly, in some embodiments, thepresent disclosure provides a method of treating pulmonary hypertension,comprising administering to a subject an effective amount of a compounddisclosed herein. The pulmonary hypertension may be WHO Group IIIpulmonary hypertension, such as pulmonary fibrosis-related pulmonaryhypertension (PH-PF) or interstitial lung disease-related pulmonaryhypertension (PH-ILD). In some embodiments, the compound is administeredby inhalation.

Other types of interstitial lung diseases include, but are not limitedto, (1) interstitial pneumonia caused by bacteria, viruses, or fungi;(2) nonspecific interstitial pneumonitis usually associated withautoimmune conditions such as rheumatoid arthritis or scleroderma; (3)hypersensitivity pneumonitis caused by inhalation of dust, mold, orother irritants; (4) cryptogenic organizing pneumonia; (5) acuteinterstitial pneumonitis; (6) desquamative interstitial pneumonitis; (7)sarcoidosis; (8) drug-induced interstitial lung disease; and (9)progressive fibrosing interstitial lung disease (PF-ILD). In someembodiments, the present disclosure provides a method of treating aninterstitial lung disease, comprising administering to a subject aneffective amount of a compound disclosed herein.

Both transforming growth factor (TGF)-beta(1) and activin-A have beenimplicated in airway remodeling in asthma (Kariyawasam, H. H., J AllergyClin Immunol., 2009, September, 124(3), pp. 454-462). In someembodiments, the present disclosure provides a method of treatingasthma, comprising administering to a subject an effective amount of acompound disclosed herein.

Chronic obstructive pulmonary disease (COPD) is a pulmonary disordercharacterized by a poorly reversible and progressive airflow limitationcaused by airway inflammation and emphysema, whereas IPF is associatedwith impaired diffusion capacity (Chilosi, M., et al., Respir. Res.,2012, 13(1), 3, pp. 1-9). Both diseases, however, demonstrate aprogressive loss of alveolar parenchyma leading to severe impairment ofrespiratory function. Fibrosis associated with emphysema is known andresearch has demonstrated TGF-β1 involvement in chronic sinus disease,pulmonary fibrosis, asthma, and COPD (Yang, Y. C., et al., Allergy,2012, 67, pp. 1193-1202). In some embodiments, the present disclosureprovides a method of treating COPD, comprising administering to asubject an effective amount of a compound disclosed herein.

Other types of lung injury that result in fibrosis includesilica-induced pneumoconiosis (silicosis), asbestos-induced pulmonaryfibrosis (asbestosis), and chemotherapeutic agent-induced pulmonaryfibrosis. In some embodiments, the present disclosure provides a methodof treating injury-induced fibrosis, comprising administering to asubject an effective amount of a compound disclosed herein.

In some embodiments, the present disclosure provides a method oftreating liver fibrosis, comprising administering to a subject aneffective amount of a compound disclosed herein. Fibrosis develops inthe liver when it is repeatedly or continuously damaged, for example, inpatients with chronic hepatitis. TGF-0 signaling participates in allstages of disease progression, from initial liver injury throughinflammation and fibrosis, to cirrhosis and cancer (Fabregat, I., etal., The FEBS J., 2016, 283(12), pp. 2219-2232).

A related condition involves fibrosis resulting from idiopathicnon-cirrhotic portal hypertension (INCPH). This disease is of uncertainetiology characterized by periportal fibrosis and involvement of smalland medium branches of the portal vein. According to Nakanuma et al.,small portal veins and skin findings are similar between patients withscleroderma and INCPH (Nakanuma, Y., Hepatol. Res., 2009, 39, pp.1023-1031). Transforming growth factor-β (TGF-β) and connective tissuegrowth factor, which are fibrosis-related and vascular endothelialgrowth factors, respectively, increase in serum, skin, and the portalvein, suggesting that these could be mechanisms of the portal veinocclusion in INCPH. Moreover, endothelial mesenchymal transition (EndMT)theory was proposed by Kitao et al. based on these findings (Kitao, A.,et al., Am. J. Pathol., 2009, 175, pp. 616-626). The increase of TGF-βin sera may act as a potent inducer of EndMT. In some embodiments, thepresent disclosure provides a method of treating INCPH, comprisingadministering to a subject an effective amount of a compound disclosedherein.

Other types of liver fibrosis include alcoholic and non-alcoholic liverfibrosis, hepatitis C-induced liver fibrosis, primary biliary cirrhosisor cholangitis, and parasite-induced liver fibrosis (schistosomiasis).In some embodiments, the present disclosure provides a method oftreating alcoholic liver fibrosis, non-alcoholic liver fibrosis,hepatitis C-induced liver fibrosis, primary biliary cirrhosis, primarybiliary cholangitis, or parasite-induced liver fibrosis(schistosomiasis), comprising administering to a subject an effectiveamount of a compound disclosed herein.

Primary biliary cholangitis (PBC) and primary sclerosing cholangitis(PSC) are two types of chronic liver disease that often lead tocirrhosis and liver failure. Liver biopsies of patients with PBC or PSCtypically reveal inflammation and fibrosis. Inhibition of integrin αvβ6,which has been shown to bind to and activate TGFβ1 on epithelial cells,suppresses biliary fibrosis in rodents. (Peng, Z-W., et al., Hepatology,2016, 63, pp. 217-232). Accordingly, inhibition of the TGF-β pathway isalso expected to suppress fibrotic processes in both PBC and PSC.Compounds of the present disclosure are expected to inhibit TGF-βsignaling in liver tissue and prevent, halt, slow or reverse theprogression of PBC and PSC. Thus, in some embodiments, the presentdisclosure provides a method of treating primary biliary cholangitis orprimary sclerosing cholangitis, comprising administering to a subject aneffect amount of a compound described herein. In some embodiments, thepresent disclosure provides a method of treating liver fibrosis,optionally in a subject that suffers from PBC or PSC, comprisingadministering to the subject an effective amount of a compound describedherein.

Fibrotic skin conditions include, but are not limited to, hypertrophicscarring, keloids, and localized or systemic sclerosis (scleroderma). Asdiscussed previously, TGF-β is a potent stimulus of connective tissueaccumulation and has been implicated in the pathogenesis of sclerodermaand other fibrotic disorders (Lakos, G., et al., Am. J. Pathol., 2004,165(1), pp. 203-217). Lakos et. al. demonstrated that Smad3 functions asa key intracellular signal transducer for profibrotic TGF-β responses innormal skin fibroblasts and found that the targeted disruption ofTGF-β/Smad3 signaling modulated skin fibrosis in the mouse model ofscleroderma. In some embodiments, the present disclosure provides amethod of treating skin fibrosis, comprising administering to a subjectan effective amount of a compound disclosed herein.

Intestinal fibrosis is a common complication of inflammatory boweldisease (IBD) and is a serious clinical problem. TGF-β has beenimplicated as a major driving factor of intestinal fibrosis. Moreover,TGF-β1 signaling contributes to stricture formation in fibrostenoticCrohn's disease by inducing insulin-like growth factor I (IGF-I) andmechano-growth factor (MGF) production in intestinal smooth muscle.(Latella, G., Rieder, F., Curr. Opin. Gastroenterol., 2017, 33(4), pp.239-245). Inhibition of TGF-β signaling could thus slow, halt or reversethe progression of fibrosis in the intestine. However, adverse sideeffects of concern to patients with IBD-such as inflammation andneoplasia-would likely result from systemic inhibition of TGF-βsignaling. One goal of the present disclosure is to develop ALK5inhibitors with high selectivity for the gastrointestinal tract andrapid clearance. In some embodiments, the present disclosure provides amethod of treating intestinal fibrosis, comprising administering to asubject an effective amount of a compound described herein, for example,by once or twice daily administration of an oral ALK5 inhibitor havingminimal systemic exposure. In some embodiments, the subject suffers frominflammatory bowel disease, such as Crohn's disease or colitis. Thedegree of therapeutic efficacy may be with respect to a startingcondition of the subject (e.g., a baseline Mayo score, baselineLichtiger score, or severity or incidence of one or more symptoms), orwith respect to a reference population (e.g., an untreated population,or a population treated with a different agent). Severity of intestinalfibrosis may be assessed using any suitable method, such as delayedenhancement MRI, ultrasound elastography, shear wave elastography,magnetization MRI, or by the direct detection of macromolecules such ascollagen. Preferably, treatment with a compound of the presentdisclosure reduces the severity of the fibrosis, such as from severefibrosis to moderate or mild fibrosis. In some embodiments, thetreatment increases intestinal tissue elasticity, reduces tissuestiffness, and/or reduces collagen levels. In some embodiments, thetreatment prevents myofibroblast accumulation, inhibits expression ofpro-fibrotic factors, and/or inhibits accumulation of fibrotic tissue.

Other types of organ-specific fibrosis or fibrotic diseases involvingthe TGF-β pathway include, but are not limited to, radiation-inducedfibrosis (various organs), bladder fibrosis, intestinal fibrosis,peritoneal sclerosis, diffuse fasciitis, Dupuytren's disease,myelofibrosis, oral submucous fibrosis, and retinal fibrosis. In someembodiments, the present disclosure provides a method of treatingradiation-induced fibrosis, bladder fibrosis, intestinal fibrosis,peritoneal sclerosis, diffuse fasciitis, Dupuytren's disease,myelofibrosis, oral submucous fibrosis, or retinal fibrosis, comprisingadministering to a subject an effective amount of a compound disclosedherein.

Although one of the goals of this disclosure is to treat fibrotic andpulmonary diseases locally or in a targeted way, the compounds describedherein may also be used to treat patients systemically. Diseases thatmay be treated systemically, include, for example, oncologic diseasessuch as glioblastoma, pancreatic cancer and hepatocellular carcinoma,breast cancer metastasized to lungs, non-small cell lung cancer, smallcell lung cancer, cystic fibrosis, and metastasis of other forms ofprimary cancer subtypes. Some of the forgoing diseases may also betreated locally as well.

Other fibrotic diseases that compounds disclosed herein may treatinclude angioedema, anti-aging, and alopecia. Alopecia includes alopeciatotalis, alopecia universalis, androgenetic alopecia, alopecia areata,diffuse alopecia, postpartum alopecia, and traction alopecia.

Other Indications

In certain aspects, the present disclosure provides a method ofreversing one or more symptoms of aging, comprising administering to asubject an ALK5 inhibitor. The method may further comprise administeringan activator of the MAPK pathway, such as oxytocin. The method may beeffective in one or more of enhancing neurogenesis in the hippocampus,reducing neuroinflammation, improving cognitive ability, reducing liveradiposity, reducing liver fibrosis, and decreasing the number of p16⁺cells. In some embodiments, a method described herein increases stemcell activity. The increase in stem cell activity may allow the subjectto generate new muscle fibers and/or to form new neurons in thehippocampus. Treatment with an ALK5 inhibitor, such as a compounddescribed herein, may prevent or slow the onset of age-related diseases,such as Alzheimer's disease. (see Mehdipour, M. et al. Aging 2018, 10,5628-5645).

Pharmaceutical Compositions

In some aspects, the present disclosure provides a pharmaceuticalcomposition. The pharmaceutical composition may comprise a compounddisclosed herein, such as a compound of Formula (I), (I′), (I-A),(I′-A), (I-B), (I′-B), (I-C), (I′-C), (I-D), (I′-D), (I-E) or (I′-E),and a pharmaceutically acceptable carrier. In some embodiments, thepharmaceutical composition is formulated for oral administration. Insome embodiments, the pharmaceutical composition is formulated forinhalation. In some embodiments, the pharmaceutical compositioncomprises a compound disclosed herein and an additional therapeuticagent. Non-limiting examples of such therapeutic agents are describedherein below.

Pharmaceutical compositions typically include at least onepharmaceutically acceptable carrier, diluent or excipient and at leastone compound of Formula (I), (I′), (I-A), (I′-A), (I-B), (I′-B), (I-C),(I′-C), (I-D), (I′-D), (I-E) or (I′-E), or a compound provided in Table1 or 2-described herein as the active agent. The active agent may beprovided in any form suitable for the particular mode of administration,such as a free base, a free acid, or a pharmaceutically acceptable salt.Additionally, the methods and pharmaceutical compositions of the presentdisclosure include the use of N-oxides, crystalline forms (e.g.,polymorphs), as well as metabolites of these compounds having similaractivity. All tautomers of the compounds described herein are includedwithin the scope of the present disclosure. Additionally, the compoundsdescribed herein encompass unsolvated as well as solvated forms withpharmaceutically acceptable solvents such as water, ethanol and thelike.

Suitable routes of administration include, but are not limited to, oral,intravenous, rectal, vaginal, aerosol, pulmonary, nasal, transmucosal,topical, transdermal, otic, ocular, and parenteral modes ofadministration. In addition, by way of example only, parenteral deliveryincludes intramuscular, subcutaneous, intravenous, intramedullaryinjections, as well as intrathecal, direct intraventricular,intraperitoneal, intralymphatic, and intranasal injections.

In certain embodiments, a compound described herein is administered in alocal rather than systemic manner, for example, via injection of thecompound directly into an organ, often in a depot preparation orsustained release formulation. In some embodiments, a long actingformulation is administered by implantation (e.g., subcutaneously orintramuscularly) or by intramuscular injection. In some embodiments, acompound described herein is provided in the form of a rapid releaseformulation, an extended release formulation, or an intermediate releaseformulation. In some embodiments, a compound described herein isprovided in the form of a nebulized formulation. In some embodiments, acompound described herein is administered locally to the lungs byinhalation.

Compounds of the present disclosure are effective over a wide dosagerange. For example, in the treatment of adult humans, dosages from 0.01to 1000 mg, 0.5 to 100 mg, 1 to 50 mg, or from 5 to 40 mg per day may beadministered to a subject in need thereof. The exact dosage will dependupon the route of administration, the form in which the compound isadministered, the subject to be treated, the body weight of the subjectto be treated, and the preference and experience of the attendingphysician.

A compound of the present disclosure may be administered in a singledose. In some embodiments, a compound of the disclosure is administeredin multiple doses, such as about once, twice, three times, four times,five times, six times, or more than six times per day. In someembodiments, dosing is about once a month, once every two weeks, once aweek, or once every other day. In some embodiments, a compound of thedisclosure and an additional therapeutic agent are administered togetherabout once per day to about 6 times per day. In some embodiments, theadministration continues for more than about 6, 10, 14, 28 days, twomonths, six months, or more than about one year. In some embodiments, adosing schedule is maintained as long as necessary. A compound of thepresent disclosure may be administered chronically on an ongoing basis,e.g., for the treatment of chronic effects.

Pharmaceutical compositions of the present disclosure typically containa therapeutically effective amount of a compound of the presentdisclosure. Those skilled in the art will recognize, however, that apharmaceutical composition may contain more than a therapeuticallyeffective amount, e.g., bulk compositions, or less than atherapeutically effective amount, e.g., individual unit doses designedfor co-administration to achieve a therapeutically effective amount.

Typically, pharmaceutical compositions of the present disclosure containfrom about 0.01 to about 95% by weight of the active agent; including,for example, from about 0.05 to about 30% by weight; and from about 0.1%to about 10% by weight of the active agent.

Any conventional carrier or excipient may be used in the pharmaceuticalcompositions of the present disclosure. The choice of a particularcarrier or excipient, or combinations of carriers or excipients, willdepend on the mode of administration being used to treat a particularpatient or type of medical condition or disease state. Additionally, thecarriers or excipients used in the pharmaceutical compositions of thisdisclosure may be commercially-available. Conventional formulationtechniques are described in Remington: The Science and Practice ofPharmacy, 20th Edition, Lippincott Williams & White, Baltimore, Maryland(2000); and H. C. Ansel et al., Pharmaceutical Dosage Forms and DrugDelivery Systems, 7th Edition, Lippincott Williams & White, Baltimore,Maryland (1999).

Representative examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, the following:sugars, such as lactose, glucose and sucrose; starches, such as cornstarch and potato starch; cellulose, such as microcrystalline celluloseand its derivatives, such as sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin;talc; excipients, such as cocoa butter and suppository waxes; oils, suchas peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil,corn oil and soybean oil; glycols, such as propylene glycol; polyols,such as glycerin, sorbitol, mannitol and polyethylene glycol; esters,such as ethyl oleate and ethyl laurate; agar; buffering agents, such asmagnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-freewater; isotonic saline; Ringer's solution; ethyl alcohol; phosphatebuffer solutions; and other non-toxic compatible substances employed inpharmaceutical compositions.

Pharmaceutical compositions are typically prepared by thoroughly andintimately mixing or blending the active agent with apharmaceutically-acceptable carrier and one or more optionalingredients. The resulting uniformly blended mixture can then be shapedor loaded into tablets, capsules, pills and the like using conventionalprocedures and equipment.

In one aspect, the pharmaceutical composition is suitable for inhaledadministration. Pharmaceutical compositions for inhaled administrationare typically in the form of an aerosol or a powder. Such compositionsare generally administered using inhaler delivery devices, such as a drypowder inhaler (DPI), a metered-dose inhaler (MDI), a nebulizer inhaler,or a similar delivery device.

In a particular embodiment, the pharmaceutical composition isadministered by inhalation using a dry powder inhaler. Such dry powderinhalers typically administer the pharmaceutical composition as afree-flowing powder that is dispersed in a patient's air-stream duringinspiration. In order to achieve a free-flowing powder composition, thetherapeutic agent is typically formulated with a suitable excipient suchas lactose, starch, mannitol, dextrose, polylactic acid (PLA),polylactide-co-glycolide (PLGA) or combinations thereof. Typically, thetherapeutic agent is micronized and combined with a suitable carrier toform a composition suitable for inhalation.

A representative pharmaceutical composition for use in a dry powderinhaler comprises lactose and a micronized form of a compound disclosedherein. Such a dry powder composition can be made, for example, bycombining dry milled lactose with the therapeutic agent and then dryblending the components. The composition is then typically loaded into adry powder dispenser, or into inhalation cartridges or capsules for usewith a dry powder delivery device.

Dry powder inhaler delivery devices suitable for administeringtherapeutic agents by inhalation are described in the art and examplesof such devices are commercially available. For example, representativedry powder inhaler delivery devices or products include Aeolizer(Novartis); Airmax (IVAX); ClickHaler (Innovata Biomed); Diskhaler(GlaxoSmithKline); Diskus/Accuhaler (GlaxoSmithKline); Ellipta(GlaxoSmithKline); Easyhaler (Orion Pharma); Eclipse (Aventis); FlowCaps(Hovione); Handihaler (Boehringer Ingelheim); Pulvinal (Chiesi);Rotahaler (GlaxoSmithKline); SkyeHaler/Certihaler (SkyePharma);Twisthaler (Schering-Plough); Turbuhaler (AstraZeneca); Ultrahaler(Aventis); and the like.

A pharmaceutical composition of the present disclosure may beadministered by inhalation using a metered-dose inhaler. Suchmetered-dose inhalers typically discharge a measured amount of atherapeutic agent using a compressed propellant gas. Accordingly,pharmaceutical compositions administered using a metered-dose inhalertypically comprise a solution or suspension of the therapeutic agent ina liquefied propellant. Any suitable liquefied propellant may beemployed, including hydrofluoroalkanes (HFAs), such as1,1,1,2-tetrafluoroethane (HFA 134a) and1,1,1,2,3,3,3-heptafluoro-n-propane, (HFA 227); and chlorofluorocarbons,such as CCl₃F. In a particular embodiment, the propellant is ahydrofluoroalkane. In some embodiments, the hydrofluoroalkaneformulation contains a co-solvent, such as ethanol or pentane, and/or asurfactant, such as sorbitan trioleate, oleic acid, lecithin, andglycerin.

A representative pharmaceutical composition for use in a metered-doseinhaler comprises from about 0.01% to about 5% by weight of a compoundof the present disclosure; from about 0% to about 20% by weight ethanol;and from about 0% to about 5% by weight surfactant; with the remainderbeing an HFA propellant. Such compositions are typically prepared byadding chilled or pressurized hydrofluoroalkane to a suitable containercontaining the therapeutic agent, ethanol (if present) and thesurfactant (if present). To prepare a suspension, the therapeutic agentis micronized and then combined with the propellant. The composition isthen loaded into an aerosol canister, which typically forms a portion ofa metered-dose inhaler device.

Metered-dose inhaler devices suitable for administering therapeuticagents by inhalation are described in the art and examples of suchdevices are commercially available. For example, representativemetered-dose inhaler devices or products include AeroBid Inhaler System(Forest Pharmaceuticals); Atrovent Inhalation Aerosol (BoehringerIngelheim); Flovent (GlaxoSmithKline); Maxair Inhaler (3M); ProventilInhaler (Schering); Serevent Inhalation Aerosol (GlaxoSmithKline); andthe like.

A pharmaceutical composition of the present disclosure may beadministered by inhalation using a nebulizer inhaler. Such nebulizerdevices typically produce a stream of high velocity air that causes thepharmaceutical composition to spray as a mist that is carried into thepatient's respiratory tract. Accordingly, when formulated for use in anebulizer inhaler, the therapeutic agent can be dissolved in a suitablecarrier to form a solution. Alternatively, the therapeutic agent can bemicronized or nanomilled and combined with a suitable carrier to form asuspension.

A representative pharmaceutical composition for use in a nebulizerinhaler comprises a solution or suspension comprising from about 0.05μg/mL to about 20 mg/mL of a compound of the present disclosure andexcipients compatible with nebulized formulations. In one embodiment,the solution has a pH of about 3 to about 8.

Nebulizer devices suitable for administering therapeutic agents byinhalation are described in the art and examples of such devices arecommercially available. For example, representative nebulizer devices orproducts include the Respimat® Softmist™ Inhalaler (BoehringerIngelheim); the AERx® Pulmonary Delivery System (Aradigm Corp.); thePARI LC Plus® Reusable Nebulizer or PARI eFlow® rapid Nebulizer System(Pari GmbH); and the like.

A pharmaceutical composition of the present disclosure may be preparedin a dosage form intended for oral administration. Suitablepharmaceutical compositions for oral administration may be in the formof capsules, tablets, pills, lozenges, cachets, dragees, powders,granules; or as a solution or a suspension in an aqueous or non-aqueousliquid; or as an oil-in-water or water-in-oil liquid emulsion; or as anelixir or syrup; and the like; each containing a predetermined amount ofa compound of the present disclosure as an active ingredient.

When intended for oral administration in a solid dosage form, thepharmaceutical compositions of the disclosure will typically comprisethe active agent and one or more pharmaceutically-acceptable carriers,such as sodium citrate or dicalcium phosphate. Optionally oralternatively, such solid dosage forms may also comprise: fillers orextenders, binders, humectants, solution retarding agents, absorptionaccelerators, wetting agents, absorbents, lubricants, coloring agents,and buffering agents. Release agents, wetting agents, coating agents,sweetening, flavoring and perfuming agents, preservatives andantioxidants can also be present in the present pharmaceuticalcompositions.

Alternative formulations may include controlled release formulations,liquid dosage forms for oral administration, transdermal patches, andparenteral formulations. Conventional excipients and methods ofpreparation of such alternative formulations are described, for example,in the reference by Remington, supra.

The following non-limiting examples illustrate representativepharmaceutical compositions of the present disclosure.

Dry Powder Composition

A micronized compound of the present disclosure (1 g) is blended withmilled lactose (25 g). This blended mixture is then loaded intoindividual blisters of a peelable blister pack in an amount sufficientto provide between about 0.1 mg to about 4 mg of the compound per dose.The contents of the blisters are administered using a dry powderinhaler.

Dry Powder Composition

A micronized compound of the present disclosure (1 g) is blended withmilled lactose (20 g) to form a bulk composition having a weight ratioof compound to milled lactose of 1:20. The blended composition is packedinto a dry powder inhalation device capable of delivering between about0.1 mg to about 4 mg of the compound per dose.

Metered-Dose Inhaler Composition

A micronized compound of the present disclosure (10 g) is dispersed in asolution prepared by dissolving lecithin (0.2 g) in demineralized water(200 mL). The resulting suspension is spray dried and then micronized toform a micronized composition comprising particles having a meandiameter less than about 1.5 m. The micronized composition is thenloaded into metered-dose inhaler cartridges containing pressurized1,1,1,2-tetrafluoroethane in an amount sufficient to provide about 0.1mg to about 4 mg of the compound per dose when administered by themetered dose inhaler.

Nebulizer Composition

A representative nebulizer composition is as follows. A compound of thepresent disclosure (2 g of free-base equivalents) is dissolved in asolution containing 80 mL reverse-osmosis water, 0.1-1% by weight ofanhydrous citric acid, and 0.5-1.5 equivalents of hydrochloric acid,followed by addition of sodium hydroxide to adjust the pH to 3.5 to 5.5.Thereafter, between 4-6% by weight of D-mannitol is added and solutionq.s. to 100 mL. The solution is then filtered through a 0.2 μm filterand stored at room temperature prior to use. The solution isadministered using a nebulizer device that provides about 0.1 mg toabout 4 mg of the compound per dose.

Kits

In certain aspects, the present disclosure provides a kit comprising oneor more unit doses of a compound or pharmaceutical composition describedherein, optionally wherein the kit further comprises instructions forusing the compound or pharmaceutical composition. In some embodiments,the kit comprises a carrier, package, or container that iscompartmentalized to receive one or more containers, such as vials,tubes, and the like, each of the container(s) comprising one of theseparate elements to be used in a method described herein. Suitablecontainers include, for example, bottles, vials, syringes, and testtubes. The containers may be formed from a variety of materials, such asglass or plastic.

The articles of manufacture provided herein may contain packagingmaterials. Packaging materials for use in packaging pharmaceuticalproducts include those found in, e.g., U.S. Pat. Nos. 5,323,907,5,052,558 and 5,033,252. Examples of pharmaceutical packaging materialsinclude, but are not limited to, blister packs, bottles, tubes,inhalers, pumps, bags, vials, containers, syringes, bottles, and anypackaging material suitable for a selected formulation and intended modeof administration and treatment. For example, the container(s) mayinclude one or more compounds described herein, optionally in acomposition or in combination with another agent as disclosed herein.The container(s) may optionally have a sterile access port (for example,the container is an intravenous solution bag or a vial having a stopperpierceable by a hypodermic injection needle). Such kits may optionallycomprise a compound with an identifying description or label orinstructions relating to its use in the methods described herein.

In some embodiments, a kit includes one or more containers, each withone or more of various materials (such as reagents, optionally inconcentrated form, and/or devices) desirable from a commercial and userstandpoint for use of a compound described herein. Nonlimiting examplesof such materials include, but are not limited to, buffers, diluents,filters, needles, syringes, carrier, package, container, vial and/ortube labels listing contents and/or instructions for use, and packageinserts with instructions for use. A set of instructions will alsotypically be included. A label is optionally on or associated with thecontainer. For example, a label is on a container when letters, numbersor other characters forming the label are attached, molded, or etchedonto the container itself, a label is associated with a container whenit is present within a receptacle or carrier that also holds thecontainer, e.g., as a package insert. In addition, a label is used toindicate that the contents are to be used for a specific therapeuticapplication. In addition, the label indicates directions for use of thecontents, such as in the methods described herein. In certainembodiments, the pharmaceutical composition is presented in a pack ordispenser device which contains one or more unit dosage forms containinga compound provided herein. The pack may contain metal or plastic foil,such as a blister pack. In some embodiments, the pack or dispenserdevice is accompanied by instructions for administration. Optionally,the pack or dispenser is accompanied with a notice associated with thecontainer in a form prescribed by a governmental agency regulating themanufacture, use, or sale of pharmaceuticals, which notice is reflectiveof approval by the agency of the form of the drug for human orveterinary administration. Such notice, for example, is the labelingapproved by the U.S. Food and Drug Administration for prescriptiondrugs, or the approved product insert. In some embodiments, compositionscontaining a compound provided herein formulated in a compatiblepharmaceutical carrier are prepared, placed in an appropriate container,and labeled for treatment of an indicated condition.

Combination Therapy

The compounds and pharmaceutical compositions of the disclosure may beused in combination with one or more therapeutic agents which act by thesame mechanism or by a different mechanism to treat a disease. The oneor more agents may be administered sequentially or simultaneously, inseparate compositions or in the same composition. Useful classes ofagents for combination therapy include, but are not limited to,compounds used to treat cardiac, kidney, pulmonary, liver, skin,immunological and oncological conditions.

In practicing any of the subject methods, an ALK5 inhibitor and a secondtherapeutic agent can be administered sequentially, wherein the twoagents are introduced into a subject at two different time points. Thetwo time points can be separated by more than 2 hours, 1 or more days, 1or more weeks, 1 or more months, or according to any intermittentregimen schedule disclosed herein.

In some embodiments, the ALK5 inhibitor and the second therapeutic agentare administered simultaneously. The two agents may form part of thesame composition, or the two agents may be provided in one or more unitdoses.

In some embodiments, the ALK5 inhibitor or the second therapeutic agentare administered parenterally, orally, inhalatively, intraperitoneally,intravenously, intraarterially, transdermally, intramuscularly,liposomally, via local delivery by catheter or stent, subcutaneously,intraadiposally, or intrathecally. As used herein, a therapeuticallyeffective amount of a combination of an ALK5 inhibitor and a secondtherapeutic agent refers to a combination of an ALK5 inhibitor and asecond therapeutic agent, wherein the combination is sufficient toaffect the intended application, including but not limited to, diseasetreatment, as defined herein. Also contemplated in the subject methodsis the use of a sub-therapeutic amount of an ALK5 inhibitor and a secondtherapeutic agent in combination for treating an intended diseasecondition. The individual components of the combination, though presentin sub-therapeutic amounts, synergistically yield an efficacious effectand/or reduced a side effect in an intended application.

The amount of the ALK5 inhibitor and the second therapeutic agentadministered may vary depending upon the intended application (in vitroor in vivo), or the subject and disease condition being treated, e.g.,the weight and age of the subject, the severity of the diseasecondition, the manner of administration and the like, which can readilybe determined by one of ordinary skill in the art.

Measuring an immune response and/or the inhibition of biological effectsof ALK5 can comprise performing an assay on a biological sample, such asa sample from a subject. Any of a variety of samples may be selected,depending on the assay. Examples of samples include, but are not limitedto blood samples (e.g. blood plasma or serum), exhaled breath condensatesamples, bronchoalveolar lavage fluid, sputum samples, urine samples,and tissue samples.

A subject being treated with an ALK5 inhibitor and a second therapeuticagent may be monitored to determine the effectiveness of treatment, andthe treatment regimen may be adjusted based on the subject'sphysiological response to treatment. For example, if inhibition of abiological effect of ALK5 inhibition is above or below a threshold, thedosing amount or frequency may be decreased or increased, respectively.Alternatively, the treatment regimen may be adjusted with respect to animmune response. The methods can further comprise continuing the therapyif the therapy is determined to be efficacious. The methods can comprisemaintaining, tapering, reducing, or stopping the administered amount ofa compound or compounds in the therapy if the therapy is determined tobe efficacious. The methods can comprise increasing the administeredamount of a compound or compounds in the therapy if it is determined notto be efficacious. Alternatively, the methods can comprise stoppingtherapy if it is determined not to be efficacious. In some embodiments,treatment with an ALK5 inhibitor and a second therapeutic agent isdiscontinued if inhibition of the biological effect is above or below athreshold, such as in a lack of response or an adverse reaction. Thebiological effect may be a change in any of a variety of physiologicalindicators.

Specific agents that may be used in combination with the compoundsdisclosed herein include, but are not limited to, OFEV® (nintedanib) andEsbriet® (pirfenidone). In some embodiments, a compound disclosed hereinis administered in combination with pirfenidone, optionally wherein thepirfenidone is administered by inhalation. In some embodiments, thepresent disclosure provides a method of treating fibrosis, such asidiopathic pulmonary fibrosis, in a subject, comprising administering tothe subject an ALK5 inhibitor, such as a compound disclosed in Table 1,and nintedanib or pirfenidone. In some embodiments, the presentdisclosure provides a method of treating cancer, such as lung cancer, ina subject, comprising administering to the subject an ALK5 inhibitor,such as a compound disclosed in Table 1, and nintedanib or pirfenidone.

In some embodiments, the present disclosure provides a method fortreating a proliferative disorder (e.g., lung cancer) in a subject inneed thereof, comprising administering to said subject an ALK5 inhibitorand an immunotherapeutic agent. TGF-β has been shown to regulatelymphocyte differentiation, suppress T cell proliferation and to enhancetumor growth. Moreover, TGF-β has been shown to prevent optimalactivation of the immune system in immunotherapy-resistant patients (seeLdffek, S. J. Oncolo. 2018, 1-9; incorporated herein by reference in itsentirety). Not wishing to be bound by any particular theory, the presentinventors expect that inhibition of ALK5 may enhance the efficacy of aparticular immunotherapy. As such, treatment with an immunotherapeuticagent, such as durvalumab or pembrolizumab, and an ALK5 inhibitor, suchas a compound of the present disclosure, is expected to improve theclinical outcome of a subject with cancer, such as a subject withnon-small cell lung cancer. The combination is expected to produce asynergistic effect. A synergistic combination is also expected for atriple combination of radiation therapy, immunotherapy, and ALK5inhibition. In addition, the ALK5 inhibitor, even when administeredlocally (e.g., to the lung by inhalation), may stimulate both local andsystemic immune responses, allowing for the treatment of primary ormetastatic tumors in tissues beyond the site of the local delivery. Forexample, an inhaled ALK5 inhibitor may be administered in combinationwith an immunotherapeutic agent to treat melanoma, renal cell carcinoma,colon cancer, or breast cancer.

In some embodiments, the ALK5 inhibitor and the immunotherapeutic agentare administered sequentially or simultaneously. In some embodiments,the ALK5 inhibitor and the immunotherapeutic agent are more effective intreating the proliferative disorder than either agent alone. In someembodiments, the ALK5 inhibitor and the immunotherapeutic agent yield asynergistic effect in treating the proliferative disorder. Thesynergistic effect may be a therapeutic effect that is greater thaneither agent used alone in comparable amounts under comparableconditions. The synergistic effect may be a therapeutic effect that isgreater than results expected by adding the effects of each agent alone.In some embodiments, the proliferative disorder is a cancer condition.In some embodiments, the cancer condition is lung cancer, such asnon-small cell lung cancer.

The term “immunotherapeutic agent” refers to any agent that induces,enhances, suppresses or otherwise modifies an immune response. Thisincludes the administration of an active agent to, or any type ofintervention or process performed on, the subject, with the objective ofmodifying an immune response. An immunotherapeutic agent may, forexample, increase or enhance the effectiveness or potency of an existingimmune response in a subject, for example, by stimulating mechanismsthat enhance the endogenous host immune response or overcomingmechanisms that suppress the endogenous host immune response.

“Immune response” refers to the action of a cell of the immune systemincluding, for example, B lymphocytes, T lymphocytes, natural killer(NK) cells, macrophages, eosinophils, mast cells, myeloid-derivedsuppressor cells, dendritic cells and neutrophils and solublemacromolecules produced by any of these cells or the liver (includingantibodies, cytokines and complement), that results in selectivetargeting, binding to, damage to, destruction of, and/or elimination ofinvading pathogens, cells or tissues infected with pathogens, cancerousor other abnormal cells, or, in cases of autoimmunity or pathologicalinflammation, normal human cells or tissues, from the body of a subject.

In one embodiment, an immunotherapeutic agent may comprise a PD-1inhibitor. In another embodiment, an immunotherapeutic agent maycomprise a CTLA-4 inhibitor. In still another embodiment, animmunotherapeutic agent may comprise a B7 inhibitor.

Exemplary PD-1 inhibitors: A PD-1 inhibitor suitable for use in thesubject methods can be selected from a variety of types of molecules.For example, the PD-1 inhibitor can be a biological or chemicalcompound, such as an organic or inorganic molecule, peptide, peptidemimetic, antibody or an antigen-binding fragment of an antibody. Someexemplary classes of agents suitable for use in the subject methods aredetailed in the sections below. A PD-1 inhibitor for use in the presentdisclosure can be any PD-1 inhibitor that is known in the art, and caninclude any entity that, upon administration to a patient, results ininhibition of the PD-1 pathway in the patient. A PD-1 inhibitor caninhibit PD-1 by any biochemical mechanism, including disruption of anyone or more of PD-1/PD-L1, PD1/PD-L2 and PD-L1/CD80 interactions.

In some embodiments, the PD-1 inhibitor is a molecule that inhibits thebinding of PD-1 to its ligand binding partners. In a specific aspect,the PD-1 ligand binding partners are PD-L1 and/or PD-L2. In anotherembodiment, a PD-1 inhibitor is a molecule that inhibits the binding ofPD-L1 to its binding partners. In a specific aspect, PD-L1 bindingpartners are PD1 and/or CD80. In another embodiment, the PD-1 inhibitoris a molecule that inhibits the binding of PD-L2 to its bindingpartners. In a specific aspect, a PD-L2 binding partner is PD1. Theinhibitor may be an antibody, an antigen binding fragment thereof, animmunoadhesin, a fusion protein or oligopeptide.

In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. Insome further embodiments, the anti-PD-1 antibody is capable ofinhibiting binding between PD-1 and PD-L1. In another embodiment, theanti-PD-1 antibody is capable of inhibiting binding between PD-1 andPD-L2. In some embodiments, the PD-1 inhibitor is an anti-PD-L1antibody. In some embodiments, the anti-PD-L1 antibody is capable ofinhibiting binding between PD-L1 and PD-1 and/or between PD-L1 and CD80.In some embodiments, the PD-1 inhibitor is an anti-PD-L2 antibody. Insome further embodiments, the anti-PD-L2 antibody is capable ofinhibiting binding between PD-1 and PD-L2. In yet another embodiment,the PD-1 inhibitor is nivolumab or pembrolizumab. In some embodiments,the PD-1 inhibitor is selected from atezolizumab, avelumab, nivolumab,pembrolizumab, durvalumab and BGB-A317.

Inhibition of the PD-1 pathway can enhance the immune response tocancerous cells in a patient. The interaction between PD-1 and PD-L1impairs T cell response as manifested by a decrease intumor-infiltrating lymphocytes (TILs) and a decrease in T-cell receptormediated proliferation, resulting in T cell anergy, exhaustion orapoptosis, and immune evasion by the cancerous cells. This immunesuppression can be reversed by inhibiting the local interaction betweenPD-L1 and PD-1 using a PD-1 inhibitor, including, for example, ananti-PD-1 and/or an anti-PD-L1 Ab. A PD-1 inhibitor may improve orrestore antitumor T-cell functions.

Anti-PD-1 antibodies suitable for use in the disclosure can be generatedusing methods well known in the art. Exemplary PD-1 inhibitors include,but are not limited to: nivolumab (BMS936558), pembrolizumab (MK-3475),pidilizumab (CT-011), AMP-224, AMP-514, BMS-936559, RG7446 (MPDL3280A),MDX-1106 (Medarex Inc.), MSB0010718C, MED14736, and HenGrui mAB005 (WO15/085847). Further PD-1 antibodies and other PD-1 inhibitors includethose described in WO 04/056875, WO 06/121168, WO 07/005874, WO08/156712, WO 09/014708, WO 09/114335, WO 09/101611, WO 10/036959, WO10/089411, WO 10/027827, WO 10/077634, WO 11/066342, WO 12/145493, WO13/019906, WO 13/181452, WO 14/022758, WO 14/100079, WO 14/206107, WO15/036394, WO 15/085847, WO 15/112900, WO 15/112805, WO 15/112800, WO15/109124, WO 15/061668, WO 15/048520, WO 15/044900, WO 15/036927, WO15/035606; U. S. Pub. No. 2015/0071910; and U.S. Pat. Nos. 7,488,802;7,521,051; 7,595,048; 7,722, 868; 7,794,710; 8,008,449; 8,354,509;8,383,796; 8,652,465; and 8,735,553; all of which are incorporatedherein by reference. Some anti-PD-1 antibodies are commerciallyavailable, for example from ABCAM (AB137132), BIOLEGEND (EH12.2H7, RMP1-14) and AFFYMETRIX EBIOSCIENCE (J105, J116, M1H4).

Exemplary CTLA-4 inhibitors: A CTLA-4 inhibitor suitable for use in thesubject methods can be selected from a variety of types of molecules.For example, the CTLA-4 inhibitor can be a biological or chemicalcompound, such as an organic or inorganic molecule, peptide, peptidemimetic, antibody or an antigen-binding fragment of an antibody. Someexemplary classes of agents suitable for use in the subject methods aredetailed in the sections below. A CTLA-4 inhibitor for use in thepresent disclosure can be any CTLA-4 inhibitor that is known in the art,and can include any entity that, upon administration to a patient,results in inhibition of the CTLA-4 pathway in the patient. A CTLA-4inhibitor can inhibit CTLA-4 by any biochemical mechanism, includingdisruption of either one or both of CTLA-4/CD80 and CTLA-4/CD86interactions.

In some embodiments, the CTLA-4 inhibitor is a molecule that inhibitsthe binding of CTLA-4 to its ligand binding partners. In a specificaspect, the CTLA-4 ligand binding partners are CD80 and/or CD86. Inanother embodiment, a CTLA-4 inhibitor is a molecule that inhibits thebinding of CD80 to its binding partners. In a specific aspect, a CD80binding partner is CTLA-4. In another embodiment, the CTLA-4 inhibitoris a molecule that inhibits the binding of CD86 to its binding partners.In a specific aspect, a CD86 binding partner is CTLA-4. The inhibitormay be an antibody, an antigen binding fragment thereof, animmunoadhesin, a fusion protein or oligopeptide.

In some embodiments, the CTLA-4 inhibitor is an anti-CTLA-4 antibody. Insome further embodiments, the anti-CTLA-4 antibody is capable ofinhibiting binding between CTLA-4 and CD80. In another embodiment, theanti-CTLA-4 antibody is capable of inhibiting binding between CTLA-4 andCD86. In some embodiments, the CTLA-4 inhibitor is an anti-CD80antibody. In some embodiments, the anti-CD80 antibody is capable ofinhibiting binding between CTLA-4 and CD80. In some embodiments, theCTLA-4 inhibitor is an anti-CD86 antibody. In some further embodiments,the anti-CD86 antibody is capable of inhibiting binding between CTLA-4and CD86. In yet another embodiment, the CTLA-4 inhibitor istremelimumab or ipilimumab.

Inhibition of the CTLA-4 pathway can enhance the immune response tocancerous cells in a patient. The interaction between CTLA-4 and one ofits natural ligands, CD80 and CD86, delivers a negative regulatorysignal to T cells. This immune suppression can be reversed by inhibitingthe local interaction between CD80 or CD86 and CTLA-4 using a CTLA-4inhibitor, including, for example, an anti-CTLA-4 Ab, anti-CD80 Ab or anantiCD86 Ab. A CTLA-4 inhibitor may improve or restore antitumor T-cellfunctions.

Anti-CTLA-4 antibodies suitable for use in the disclosure can begenerated using methods well known in the art. Exemplary CTLA-4inhibitors include but are not limited to tremelimumab and ipilimumab(also known as 10D1 or MDX-010). Further CTLA-4 antibodies and otherCTLA-4 inhibitors include those described in WO 98/042752, WO 00/037504,WO 01/014424 and WO 04/035607; U. S. Pub. Nos. 2002/0039581, 2002/086014and 2005/0201994; U.S. Pat. Nos. 5,811,097; 5,855,887; 5,977,318;6,051,227; 6,207,156; 6,682,736; 6,984,720; 7,109,003; 7,132,281;7,605,238; 8,143,379; 8,318,916; 8,435,516; 8,784,815; and 8,883,984; EPPat. No. 1212422; Hurwitz et al., PNAS 1998, 95(17): 10067-10071;Camacho et al., J Clin Oncology 2004, 22(145): abstract no. 2505(antibody CP675206); and Mokyr, et al., Cancer Research 1998,58:5301-5304; each of which is incorporated herein by reference.

Also provided herein is a pharmaceutical composition comprising acompound of the disclosure or a pharmaceutically acceptable salt thereofand one or more other therapeutic agents. The therapeutic agent may beselected from the classes of agents specified above and from the listsof specific agents described above. In some embodiments, thepharmaceutical composition is suitable for delivery to the lungs. Insome embodiments, the pharmaceutical composition is suitable for inhaledor nebulized administration. In some embodiments, the pharmaceuticalcomposition is a dry powder or a liquid composition.

Further, in a method aspect, the disclosure provides a method oftreating a disease or disorder in a mammal comprising administering tothe mammal a compound of the disclosure or a pharmaceutically acceptablesalt thereof and one or more other therapeutic agents.

When used in combination therapy, the agents may be formulated in asingle pharmaceutical composition, or the agents may be provided inseparate compositions that are administered simultaneously or atseparate times, by the same or by different routes of administration.Such compositions can be packaged separately or may be packaged togetheras a kit. The two or more therapeutic agents in the kit may beadministered by the same route of administration or by different routesof administration.

EXAMPLES

The following examples are given for the purpose of illustrating variousembodiments of the disclosure and are not meant to limit the presentdisclosure in any fashion. The present examples, along with the methodsand compositions described herein, are presently representative ofpreferred embodiments, are exemplary, and are not intended aslimitations on the scope of the disclosure. Changes therein and otheruses which are encompassed within the spirit of the disclosure asdefined by the scope of the claims will occur to those skilled in theart.

The following abbreviations have the following meanings unless otherwiseindicated and any other abbreviations used herein and not defined havetheir standard, generally accepted meaning:

-   -   AcOH=acetic acid    -   AcONa=sodium acetate    -   ACN=acetonitrile    -   Atm=atmosphere    -   BINAP=(±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene    -   Boc₂O=di-tert-butyl dicarbonate    -   (Bpin)₂=bis(pinacolato)diboron    -   BrettPhos=2-(dicyclohexylphosphino)3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl    -   BrettPhos Pd G4=N-substituted 2-aminobiphenylpalladium        methanesulfonate precatalyst    -   BSA=bovine serum albumin, Fraction V    -   Cp*RuCl(PPh₃)₂=pentamethylcyclopentadienylbis        (triphenylphosphine)ruthenium(II) chloride    -   d=day(s)    -   DAST=diethylaminosulfur trifluoride    -   DCC=N,N′-dicyclohexylcarbodiimide    -   DCE=1,2-dichloroethane    -   DCM=dichloromethane or methylene chloride    -   DHP=dihydropyran    -   DIAD=diisopropyl azodicarboxylate    -   DIBAH=diisobutylaluminium hydride    -   DIPEA or DIEA=N,N-diisopropylethylamine    -   DMA or DMAc=dimethylacetamide    -   DMAP=4-dimethylaminopyridine    -   DMEDA=1,2-bis(methylamino)ethane    -   DMF=N,N-dimethylformamide    -   DMP=Dess-Martin periodinane    -   DMSO=dimethyl sulfoxide    -   DPPA=diphenylphosphoryl azide    -   DPPF=1,1′-bis(diphenylphosphino)ferrocene    -   DTT=dithiothreitol    -   EDCI=N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide        hydrochloride    -   EDTA=ethylenediaminetetraacetic acid    -   EGTA=ethylene glycol-bis(O-aminoethyl        ether)-N,N,N′,N′-tetraacetic acid    -   EtOH=ethanol    -   EtOAc or EA=ethyl acetate    -   g=gram(s)    -   h=hour(s)    -   HATU=N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium        hexafluorophosphate    -   HEPES=4-(2-hyrdroxyethyl)-1-piperazine ethanesulfonic acid    -   HOBT=hydroxybenzotriazole    -   i-PrOBPin=2-isopropyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane    -   IPA or i-PrOH=isopropyl alcohol    -   KHMDS=potassium bis(trimethylsilyl)amide    -   LAH=lithium aluminum hydride    -   LDA=lithium diisopropylamide    -   LiHDMS=hexamethyldisilazane lithium salt    -   m-CPBA=meta-chloroperoxybenzoic acid    -   MeCN=acetonitrile    -   MeOH=methanol    -   min=minute(s)    -   MTBE=methyl tert-butyl ether    -   NaHDMS=hexamethyldisilazane sodium salt    -   NBS=N-bromosuccinimide    -   n-BuLi=n-butyl lithium    -   Pd(dppf)Cl₂=[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(II)    -   Pd(OAc)₂=palladium(II) acetate    -   Pd(PPh₃)₄=tetrakis(triphenylphosphine)palladium(0)    -   Pd/C=palladium on activated carbon, 10% loading    -   Pd₂(dba)₃=tris(dibenzylideneacetone)dipalladium(0)    -   PE=petroleum ether    -   PhN₂=benzene diazonium ion    -   PTSA=p-toluenesulfonic acid    -   RT, rt, or r.t.=room temperature    -   RuPhos=2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl    -   RuPhos Pd        G2=chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)        [2-(2′-amino-1,1′-biphenyl)]palladium(II)    -   RuPhos Pd G4=ligand for Buchwald₄*h generation Palladacycle    -   SEMCl=2-(trimethylsilyl)ethoxymethyl chloride    -   SiO₂=silicon dioxide or silica    -   SPhos Pd G3=(2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl)        [2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate    -   STAB=sodium triacetoxyborohydride    -   TBAB=tetrabutylammonium bromide    -   TBSCl=tert-butyldimethylchlorosilane    -   t-BuOK=potassium tert-butoxide    -   t-BuONa=sodium tert-butoxide    -   TEA, Et₃N=triethylamine    -   TFA=trifluoroacetic acid    -   TFAA=trifluoroacetic anhydride    -   THF=tetrahydrofuran    -   TMS=tetramethylsilane    -   TosC1=p-toluenesulfonyl chloride    -   Tris-HCl=tris(hydroxymethyl)aminomethane hydrochloride    -   Tween-20=polyoxyethylene sorbitan monolaurate    -   Xantphos=4,5-bis(diphenylphosphino)-9,9-dimethylxanthene    -   XPhos Pd G4=Buchwald 4^(th) generation palladacycle

Unless noted otherwise, all materials, such as reagents, startingmaterials and solvents, were purchased from commercial suppliers, suchas Sigma-Aldrich, Fluka Riedel-de Haen, and the like, and were usedwithout further purification.

Reactions were run under nitrogen atmosphere, unless noted otherwise.The progress of reactions was monitored by thin layer chromatography(TLC), analytical high performance liquid chromatography (anal. HPLC),and mass spectrometry, the details of which are given in specificexamples.

Reactions were worked up as described specifically in each preparation;commonly, reaction mixtures were purified by extraction and otherpurification methods such as temperature- and solvent-dependentcrystallization, and precipitation. In addition, reaction mixtures wereroutinely purified by preparative HPLC, typically using Microsorb C18and Microsorb BDS column packings and conventional eluents. Progress ofreactions was typically monitored by liquid chromatography massspectrometry (LCMS). Characterization of isomers was typically done byNuclear Overhauser effect spectroscopy (NOE). Characterization ofreaction products was routinely carried out by mass spectrometry and/or¹H-NMR spectroscopy. For NMR measurement, samples were dissolved indeuterated solvent (CD₃OD, CDCl₃, or DMSO-d₆), and ¹H-NMR spectra wereacquired with a Varian Gemini 2000 instrument (400 MHz) under standardobservation conditions. Mass spectrometric identification of compoundswas typically conducted using an electrospray ionization method (ESMS)with an Applied Biosystems (Foster City, CA) model API 150 EX instrumentor an Agilent (Palo Alto, CA) model 1200 LC/MSD instrument.

Example 1: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(475)

Step A: Preparation of methyl 6-methylpicolinimidate (1-2). To asolution of 1-1 (50.0 g, 423.3 mmol) in MeOH (300 ml) was added 25%NaOMe in MeOH (100 ml, 465 mmol) at 0° C. Then the reaction was stirredat 25° C. for 16 h. The reaction mixture was concentrated in vacuum andthe crude residue was dissolved in EtOAc. Brine was added to thesolution and white solid precipitation was observed. The suspension wasfiltered and the organic layer of filtrate was separated. The organicphase was washed with brine, dried over sodium sulfate, filtered andconcentrated to give the crude residue 1-2 (58.0 g). The crude productwas used directly in the next step without further purification. [M+H]⁺calcd for C₈H₁₀N₂O 150.18, found 151.05.

Step B: Preparation of 6-methylpicolinimidamide hydrogen chloride (1-3).To a solution of 1-2 (58.0 g, 386.6 mmol) in EtOH (250 ml) and H₂O (25ml) was added NH₄Cl (22.9 g, 425.02 mmol) and the reaction was stirredat 80° C. for 16 h. The reaction was concentrated to dryness. The cruderesidue was triturated in EtOAc and then filtered to remove non-polarimpurities. The solid precipitate was treated with excess EtOH andfiltered. The filtrate was concentrated to obtain an HCl salt of thedesired product 1-3 (55.0 g). The crude product was used directly in thenext step without further purification. [M+H]⁺ calcd for C₇H₉N₃ 135.17,found 136.10.

Step C: Preparation of 2-(6-methylpyridin-2-yl)pyrimidin-4(3H)-one(1-5). To a solution of 1-3 (55.0 g, 407.4 mmol) and 1-4 (46.4 ml, 427.7mmol) in EtOH (385 ml) was added K₂CO₃ (112.4 g, 814.8 mmol). Thereaction was stirred at 80° C. for 16 h. The reaction was cooled to 25°C., filtered through a pad of Celite and concentrated to give crudeproduct. The crude product was acidified with 4N HCl and triturated withEtOAc, then filtered to give an HCl salt of 1-5 (65.0 g). The crudeproduct was used directly in the next step without further purification.[M+H]⁺ calcd for C₁₀H₉N₃O 187.20, found 187.17.

Step D: Preparation of 4-chloro-2-(6-methylpyridin-2-yl)pyrimidine(1-6). To a solution of 1-5 (10.0 g, 53.41 mmol, HCl salt) in1,4-dioxane (70 ml) was added POCl₃ (15.0 ml, 160.25 mmol). The reactionwas stirred at 90° C. for 16 h, then concentrated to dryness. The cruderesidue was poured onto ice-cold water. The pH was adjusted to 8 bysaturated NaHCO₃. The mixture was then extracted with EtOAc. The organicphase was dried over sodium sulfate and concentrated to give crudeproduct. The crude residue was triturated in pentane and filtered togive 1-6 (5.0 g). The crude product was used directly in the next stepwithout further purification. [M+H]⁺ calcd for C₁₀H₈ClN₃ 205.65, found206.09.

Step E: Preparation of2-chloro-N-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidin-4-amine(1-8). To a solution of 1-6 (3.0 g, 14.63 mmol) and 1-7 (2.26 g, 17.56mmol) in DMF (15 ml) was added solid crushed NaOH (1.17 g, 29.26 mmol)at 0° C. The reaction was stirred at 25° C. for 16 h. The reaction waspoured onto ice-cold water and the solid precipitated. The solid wascollected by filtration and dried. The crude residue was triturated withether and ACN to give 1-8 (3.5 g). The crude product was used directlyin the next step without further purification. [M+H]⁺ calcd forC₁₄H₁₁ClN₆ 298.73, found 298.94.

Step F: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(475). To a solution of 1-8 (250 mg, 0.84 mmol) in 1,4-dioxane (10 ml)was added 1-9 (298 mg, 1.67 mmol), Pd₂(dba)₃ (77 mg, 0.08 mmol),Xantphos (48 mg, 0.08 mmol), Cs₂CO₃ (545 mg, 1.67 mmol) and theresulting mixture was heated at 100° C. for 6 h under N₂. The reactionmixture was concentrated in vacuum. The residue was purified bypreparative HPLC chromatography using a gradient (25 to 55%) ofacetonitrile in water with 0.05% ammonia hydroxide to obtain the titlecompound as a white solid (62 mg). [M+H]⁺ calcd for C₂₄H₂₄N₈O 440.51,found 441.4.

Example 2: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4-diamine(418)

A vial of 1-8 (20 mg, 0.067 mmol), 2-1 (30 mg, 0.100 mmol), cesiumcarbonate (32.7 mg, 0.100 mmol), BrettPhos (3.59 mg, 6.69 μmol), andBrettPhos Pd G4 (6.16 mg, 6.69 μmol) in degassed 1,4-dioxane (1 ml) washeated at 110° C. for 16 h. The reaction mixture was concentrated invacuum. TFA (1 ml) and DCM (1 ml) were added to the residue and stirredat 25° C. for 2 h. TFA was removed in vacuum and the residue waspurified by preparative HPLC chromatography using a gradient (2 to 30%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (27.2 mg). [M+H]⁺ calcd for C₂₄H₂₅N₉ 439.53,found 440.2.

Example 3: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(5-(piperazin-1-yl)pyridin-2-yl)pyrimidine-2,4-diamine(121)

A vial of 1-8 (20 mg, 0.067 mmol), 3-1 (30 mg, 0.100 mmol), cesiumcarbonate (32.7 mg, 0.100 mmol), BrettPhos (3.59 mg, 6.69 μmol), andBrettPhos Pd G4 (6.16 mg, 6.69 μmol) in degassed 1,4-dioxane (1 ml) washeated at 110° C. for 16 h. The reaction mixture was concentrated invacuum. TFA (1 ml) and DCM (1 ml) were added to the residue and stirredat 25° C. for 2 h. TFA was removed in vacuum and the residue waspurified by preparative HPLC chromatography using a gradient (2 to 30%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (28.6 mg). [M+H]⁺ calcd for C₂₃H₂₄N₁₀ 440.52,found 441.3.

Example 4: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(2-(piperazin-1-yl)pyrimidin-5-yl)pyrimidine-2,4-diamine(86)

Step A: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine (4-1).To a vial of 1-8 (238 mg, 0.797 mmol) was added ammonia solution (7N inMeOH) (4 ml, 28.0 mmol). The resulting suspension was heated at 80° C.for 16 h. The reaction mixture was concentrated in vacuum. The cruderesidue was purified by silica-gel chromatography using a gradient (0 to15%) of MeOH in DCM to give 4-1 (108 mg) as a brown solid. [M+H]⁺ calcdfor C₁₃H₁₃N₇ 279.31, found 280.0.

Step B: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(2-(piperazin-1-yl)pyrimidin-5-yl)pyrimidine-2,4-diamine(86). A vial of 4-1 (24.41 mg, 0.087 mmol), 4-2 (20 mg, 0.058 mmol),sodium tert-butoxide (11.20 mg, 0.117 mmol), BrettPhos (6.26 mg, 0.012mmol), and BrettPhos Pd G4 (10.73 mg, 0.012 mmol) in degassed1,4-dioxane (1 ml) was heated at 110° C. for 1 h. The reaction mixturewas concentrated in vacuum. TFA (1 ml) was added to the residue andheated at 50° C. for 2 h. TFA was removed in vacuum and the residue waspurified by preparative HPLC chromatography using a gradient (2 to 60%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (17.2 mg). [M+H]⁺ calcd for C₂₂H₂₃N₁ 441.50,found 442.2.

Example 5: Synthesis of(R)-(4-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazin-2-yl)methanol(403)

Step A: Preparation of tert-butyl(R)-2-(hydroxymethyl)-4-(4-nitrophenyl)piperazine-1-carboxylate (5-3). Asolution of 5-1 (98 mg, 0.694 mmol), 5-2 (150 mg, 0.694 mmol), and DIPEA(1.21 ml, 6.94 mmol) in DMF (3 ml) was heated at 110° C. for 16 h. Thereaction was concentrated in vacuum. The crude residue was purified bysilica-gel chromatography using a gradient (0 to 15%) of MeOH in DCM togive 5-3 (157 mg) as an orange oil. [M+H]⁺ calcd for C₁₆H₂₃N₃O₅ 337.38,found 338.4.

Step B: Preparation of tert-butyl(R)-4-(4-aminophenyl)-2-(hydroxymethyl)piperazine-1-carboxylate (5-4). Asolution of 5-3 (157 mg, 0.465 mmol) was taken up in ethanol (3 ml), andPd/C (9.90 mg, 9.31 μmol) was added and the reaction mixture was purgedunder an atmosphere of H₂ and stirred at 25° C. for 16 h. The reactionmixture was filtered through a pad of Celite and concentrated in vacuumand the crude residue 5-4 (140 mg) was used directly in the next step.[M+H]⁺ calcd for C₁₆H₂₅N₃O₃ 307.40, found 308.0.

Step C: Preparation of(R)-(4-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazin-2-yl)methanol(403). A vial of 1-8 (25 mg, 0.084 mmol), 5-4 (25.7 mg, 0.084 mmol),cesium carbonate (35.4 mg, 0.109 mmol), BrettPhos (2.25 mg, 4.18 μmol),and BrettPhos Pd G4 (3.85 mg, 4.18 μmol) in degassed 1,4-dioxane (1.5ml) was heated to 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (226 μl) and DCM (118 μl) were added to theresidue and stirred at 45° C. for 30 min. TFA was removed in vacuum andthe residue was purified by preparative HPLC chromatography using agradient (2 to 60%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (10.2 mg). [M+H]⁺ calcdfor C₂₅H₂₇N₉O 469.55, found 470.1.

Example 6: Synthesis ofN2-(4-((azetidin-3-ylmethyl)amino)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(537)

Step A: Preparation of tert-butyl3-(((4-aminophenyl)amino)methyl)azetidine-1-carboxylate (6-2). Asolution of 5-1 (65 mg, 0.461 mmol), 6-1 (86 mg, 0.461 mmol), and DIPEA(805 μl, 4.61 mmol) in DMF (1 ml) was heated at 110° C. for 16 h. Thereaction was concentrated in vacuum. The crude product was purified bysilica-gel chromatography using a gradient (0 to 60%) of EtOAc inHexanes to give tert-butyl3-(((4-nitrophenyl)amino)methyl)azetidine-1-carboxylate (53 mg, 37%yield) as an orange oil. [M+H]⁺ calcd for C₁₅H₂₁N₃O₄ 307.35, found308.6. The orange oil (53 mg, 0.172 mmol) was taken up in ethanol (3ml), and Pd/C (2.42 mg, 0.017 mmol) was added and the reaction mixturewas purged under an atmosphere of H₂ and stirred at 25° C. for 16 h. Thereaction mixture was filtered through a pad of Celite and concentratedin vacuum to provide 6-2 as a crude residue (44 mg, 92% yield), useddirectly in the next step without further purification. [M+H]⁺ calcd forC₁₅H₂₃N₃O₂ 277.37, found 277.2.

Step B: Preparation ofN2-(4-((azetidin-3-ylmethyl)amino)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(537). A vial of 1-8 (48 mg, 0.162 mmol), 6-2 (44 mg, 0.159 mmol),cesium carbonate (211 mg, 0.647 mmol), BrettPhos (35 mg, 0.065 mmol),and BrettPhos Pd G4 (60 mg, 0.065 mmol) in degassed 1,4-dioxane (1.62ml) was heated at 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (1.24 ml) and DCM (1.04 ml) were added tothe residue and stirred at 45° C. for 30 min. TFA was removed in vacuumand the residue was purified by preparative HPLC chromatography using agradient (5 to 75%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (10.7 mg). [M+H]⁺ calcdfor C₂₄H₂₅N₉ 439.53, found 440.1.

Example 7: Synthesis ofN2-(4-((azetidin-3-ylmethyl)(methyl)amino)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(202)

Step A: Preparation of tert-butyl3-(((4-aminophenyl)(methyl)amino)methyl)azetidine-1-carboxylate (7-2). Asolution of 5-1 (65 mg, 0.461 mmol), 7-1 (92 mg, 0.461 mmol), and DIPEA(805 μl, 4.61 mmol) in DMF (1 ml) was heated at 110° C. for 16 h. Thereaction was concentrated in vacuum. The crude product was purified bysilica-gel chromatography using a gradient (0 to 60%) of EtOAc inHexanes to give tert-butyl3-((methyl(4-nitrophenyl)amino)methyl)azetidine-1-carboxylate (118 mg,80% yield) as an orange oil. [M+H]⁺ calcd for C₁₆H₂₃N₃O₄ 321.38, found322.2. The orange oil (118 mg, 0.367 mmol) was taken up in ethanol (3ml), and Pd/C (2.42 mg, 0.017 mmol) was added and the reaction mixturewas purged under an atmosphere of H₂ and stirred at 25° C. for 16 h. Thereaction mixture was filtered through a pad of Celite and concentratedin vacuum, and the crude residue 7-2 (113 mg) was used directly in thenext step. [M+H]⁺ calcd for C₁₆H₂₅N₃O₂ 291.40, found 291.2.

Step B: Preparation ofN2-(4-((azetidin-3-ylmethyl)(methyl)amino)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(202). A vial of 1-8 (97 mg, 0.323 mmol), 7-2 (113 mg, 0.159 mmol),cesium carbonate (211 mg, 0.647 mmol), BrettPhos (35 mg, 0.065 mmol),and BrettPhos Pd G4 (60 mg, 0.065 mmol) in degassed 1,4-dioxane (1.62ml) was heated at 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (1.24 ml) and DCM (1.04 ml) were added tothe residue and heated at 45° C. for 30 min. TFA was removed in vacuumand the residue was purified by preparative HPLC chromatography using agradient (5 to 75%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (45.5 mg). [M+H]⁺ calcdfor C₂₅H₂₇N₉ 453.55, found 454.2.

Example 8: Synthesis of(R)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(pyrrolidin-3-ylamino)phenyl)pyrimidine-2,4-diamine(278)

Step A: Preparation of tert-butyl(R)-3-((4-aminophenyl)amino)pyrrolidine-1-carboxylate (8-2). A solutionof 5-1 (100 mg, 0.709 mmol), 8-1 (132 mg, 0.709 mmol), and DIPEA (1.24ml, 7.09 mmol) in DMF (3 ml) was heated at 110° C. for 16 h. Thereaction was concentrated in vacuum. The crude product was purified bysilica-gel chromatography using a gradient (0 to 60%) of EtOAc inHexanes to give tert-butyl(R)-3-((4-nitrophenyl)amino)pyrrolidine-1-carboxylate (130 mg, 60%yield) as a yellow oil. [M+H]⁺ calcd for C₁₅H₂₁N₃O₄ 307.35, found 308.5.The yellow oil (130 mg, 0.423 mmol) was taken up in ethanol (3 mL), Pd/C(4.55 mg, 0.032 mmol) was added and the reaction mixture was purgedunder an atmosphere of H₂ and stirred at 25° C. for 16 h. The reactionmixture was filtered through a pad of Celite and concentrated in vacuumto afford the crude residue 8-2 (100 mg), used directly in the next stepwithout further purification. [M+H]⁺ calcd for C₁₅H₂₃N₃O₂ 277.37, found277.3.

Step B: Preparation of(R)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(pyrrolidin-3-ylamino)phenyl)pyrimidine-2,4-diamine(278). A vial of 1-8 (45 mg, 0.151 mmol), 8-2 (50.1 mg, 0.181 mmol),cesium carbonate (98 mg, 0.301 mmol), BrettPhos (8.09 mg, 0.015 mmol),and BrettPhos Pd G4 (13.87 mg, 0.015 mmol) in degassed 1,4-dioxane (4ml) was heated at 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (580 μl) and DCM (485 μl) were added to theresidue and the resulting mixture heated at 45° C. for 30 min. TFA wasremoved in vacuum and the residue was purified by preparative HPLCchromatography using a gradient (5 to 75%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(30.7 mg). [M+H]⁺ calcd for C₂₄H₂₅N₉ 439.53, found 440.1.

Example 9: Synthesis of4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzenesulfonamide(108)

A vial of 1-8 (30 mg, 0.100 mmol), 9-1 (21 mg, 0.121 mmol), cesiumcarbonate (42.5 mg, 0.131 mmol), BrettPhos (2.70 mg, 5.02 μmol), andBrettPhos Pd G4 (4.62 mg, 5.02 μmol) in degassed 1,4-dioxane (1 ml) washeated at 110° C. for 16 h. The reaction mixture was concentrated invacuum and the residue was purified by preparative HPLC chromatographyusing a gradient (5 to 75%) of acetonitrile in water with 0.05%trifluoroacetic acid to yield a TFA salt of the title compound (12.7mg). [M+H]⁺ calcd for C₂₀H₁₈N₈O₂S 434.48, found 435.0.

Example 10: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(5-morpholinothiazol-2-yl)pyrimidine-2,4-diamine(555)

A vial of 1-8 (20 mg, 0.067 mmol), 10-1 (24.8 mg, 0.134 mmol), cesiumcarbonate (43.6 mg, 0.134 mmol), BrettPhos (7.19 mg, 0.013 mmol), andBrettPhos Pd G4 (12.33 mg, 0.013 mmol) in degassed 1,4-dioxane (1 ml)was heated at 110° C. for 16 h. The reaction mixture was concentrated invacuum and the residue was purified by preparative HPLC chromatographyusing a gradient (5 to 65%) of acetonitrile in water with 0.05%trifluoroacetic acid to yield a TFA salt of the title compound (7.7 mg).[M+H]⁺ calcd for C₂₁H₂₁N₉OS 447.52, found 448.2.

Example 11: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(pyrrolidin-3-ylmethyl)phenyl)pyrimidine-2,4-diamine(489)

Step A: Preparation of tert-butyl(E)-3-(4-bromobenzylidene)pyrrolidine-1-carboxylate (11-3). To asolution of 11-1 (2.0 g, 10.8 mmol) in DMF (40 ml) was added NaH (561mg, 14.0 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 30min, then 11-2 (6.1 g, 11.9 mmol) in DMF (10 ml) was added. The reactionmixture was heated at 65° C. for 12 h. The mixture was poured into H₂O(400 mL), the white suspension was extracted with EA (200 mL×3), thecombined organic phase was washed with sat. NaCl (200 mL×3), dried oversolid Na₂SO₄, and concentrated under reduced pressure. The residue waspurified by silica-gel column chromatography using a gradient (0 to 5%)of EtOAc in PE to give 11-3 (800 mg) as colorless oil.

Step B: Preparation of tert-butyl(E)-3-(4-((diphenylmethylene)amino)benzylidene)pyrrolidine-1-carboxylate(11-5). To a mixture of 11-3 (800 mg, 2.36 mmol), 11-4 (472 mg, 3.60mmol) and tBuONa (364 mg, 3.78 mmol) in toluene (15 ml) was addedPd₂(dba)₃ (65 mg, 0.071 mmol) and BINAP (147 mg, 0.236 mmol) under N₂atmosphere. The reaction mixture was degassed with N₂ several times,then heated at 90° C. for 4 h. The reaction mixture was concentratedunder reduced pressure. The residue was purified by silica-gel columnchromatography using a gradient (0 to 10%) of EtOAc in PE to give 11-5(800 mg) as yellow oil.

Step C: Preparation of tert-butyl(E)-3-(4-aminobenzylidene)pyrrolidine-1-carboxylate (11-6). To a mixtureof 11-5 (800 mg, 1.31 mmol) in MeOH (10 ml) under N₂ atmosphere wasadded HONH₂—HCl (200 mg, 2.89 mmol) and NaOAc (323 mg, 3.94 mmol). Thereaction mixture was heated at 50° C. for 12 h. The reaction mixture wasdiluted with H₂O (10 ml) and the product extracted in EtOAc (10 ml×3),the combined organic layer was dried over Na₂SO₄ and concentrated underreduced pressure. The residue was purified by preparative HPLCchromatography using a gradient (30 to 60%) of acetonitrile in waterwith 0.225% formic acid to give to give 11-6 (150 mg) as light yellowoil. [M+H]⁺ calcd for C₁₆H₂₂N₂O₂ 274.36, found 275.2.

Step D: Preparation of tert-butyl3-(4-aminobenzyl)pyrrolidine-1-carboxylate (11-7). To a solution of 11-6(250 mg, 0.911 mmol) in MeOH (15 ml) was added Pd/C (30 mg, 10%). Thereaction mixture was degassed with H₂ several times, then stirred at 20°C. for 1 h. The reaction mixture was filtered through a pad of Celiteand the filtrate was concentrated under reduced pressure. The residuewas purified by preparative HPLC chromatography using a gradient (23 to53%) of acetonitrile in water with 0.225% formic acid to give 11-7 (100mg) as a yellow solid. [M+Na]⁺ calcd for C₁₆H₂₄N₂O₂ 299.37, found 299.2.

Step E: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(pyrrolidin-3-ylmethyl)phenyl)pyrimidine-2,4-diamine(489). A vial of 1-8 (30 mg, 0.100 mmol), 11-7 (33.3 mg, 0.121 mmol),cesium carbonate (65.4 mg, 0.201 mmol), Xantphos (11.62 mg, 0.020 mmol),and Pd(OAc)₂ (4.51 mg, 0.020 mmol) in degassed 1,4-dioxane (1 ml) washeated at 110° C. for 16 h. The reaction mixture was concentrated invacuum and the residue was purified by preparative HPLC chromatographyusing a gradient (2 to 50%) of acetonitrile in water with 0.05%trifluoroacetic acid to yield a TFA salt of the title compound (24.9mg). [M+H]⁺ calcd for C₂₅H₂₆N₈ 438.54, found 439.2.

Example 12: Synthesis ofN2-(3-isopropyl-2,3,4,5-tetrahydro-1H-benzo[d]azepin-7-yl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(223)

Step A: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(2,3,4,5-tetrahydro-1H-benzo[d]azepin-7-yl)pyrimidine-2,4-diamine(12-2). A vial of 1-8 (35 mg, 0.117 mmol), 12-1 (46.1 mg, 0.176 mmol),cesium carbonate (76 mg, 0.234 mmol), BrettPhos (12.58 mg, 0.023 mmol),and BrettPhos Pd G4 (21.57 mg, 0.023 mmol) in degassed 1,4-dioxane (586μl) was heated at 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (468 μl) and DCM (348 μl) were added to theresidue and heated at 45° C. for 30 min. TFA was removed in vacuum andthe residue was purified by preparative HPLC chromatography using agradient (2 to 30%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (76 mg). [M+H]⁺ calcd forC₂₄H₂₄N₈ 424.51, found 425.1.

Step B: Preparation ofN2-(3-isopropyl-2,3,4,5-tetrahydro-1H-benzo[d]azepin-7-yl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(223). A vial of 12-2 (18 mg, 0.028 mmol), 2-bromopropane (10.18 mg,0.083 mmol), and DIEA (48.2 μl, 0.276 mmol) in DMF (276 μl) was heatedat 110° C. for 8 h. The reaction mixture was concentrated in vacuum andthe residue was purified by preparative HPLC chromatography using agradient (2 to 60%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (2.3 mg). [M+H]⁺ calcdfor C₂₇H₃₀N₈ 466.59, found 467.3.

Example 13: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(1,2,3,4-tetrahydroisoquinolin-7-yl)pyrimidine-2,4-diamine(175)

A vial of 1-8 (25 mg, 0.075 mmol), 13-1 (28 mg, 0.112 mmol), cesiumcarbonate (48.6 mg, 0.149 mmol), BrettPhos (8.01 mg, 0.015 mmol), andBrettPhos Pd G4 (13.73 mg, 0.015 mmol) in degassed 1,4-dioxane (1 ml)was heated at 110° C. for 16 h. The reaction mixture was concentrated invacuum. TFA (1 ml) and DCM (1 ml) were added to the residue and stirredat 25° C. for 1 h. TFA was removed in vacuum and the residue waspurified by preparative HPLC chromatography using a gradient (5 to 65%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (16.7 mg). [M+H]⁺ calcd for C₂₃H₂₂N₈ 410.49,found 411.1.

Example 14: Synthesis ofN2-(4-(3,8-diazabicyclo[3.2.1]octan-8-yl)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(221)

Step A: Preparation of tert-butyl8-(4-aminophenyl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate (14-2). Asolution of 5-1 (75 mg, 0.532 mmol), 14-1 (124 mg, 0.585 mmol), andpotassium carbonate (147 mg, 1.063 mmol) in DMF (1.77 ml) was heated at80° C. for 16 h. The reaction was concentrated in vacuum. The crudeproduct was purified by silica-gel chromatography using a gradient (0 to15%) of MeOH in DCM with 2.5% triethylamine to give tert-butyl8-(4-nitrophenyl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate (68 mg,39% yield) as an orange solid. [M+H]⁺ calcd for C₁₇H₂₃N₃O₄ 333.39, found334.0. The orange solid (68 mg, 0.204 mmol) was taken up in ethanol(1.61 ml), and Pd/C (51.4 mg, 0.097 mmol) was added and the reactionmixture was purged under an atmosphere of H₂ and stirred at 25° C. for16 h. The reaction mixture was filtered through a pad of Celite andconcentrated in vacuum to give the crude residue 14-2 (55 mg), useddirectly in the next step. [M+H]⁺ calcd for C₁₅H₂₅N₃O₂ 303.41, found304.0.

Step B: Preparation ofN2-(4-(3,8-diazabicyclo[3.2.1]octan-8-yl)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(221). A vial of 1-8 (25 mg, 0.084 mmol), 14-2 (25.4 mg, 0.084 mmol),cesium carbonate (35.4 mg, 0.109 mmol), BrettPhos (2.25 mg, 4.18 μmol),and BrettPhos Pd G4 (3.85 mg, 4.18 μmol) in degassed 1,4-dioxane (1.5ml) was heated at 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (226 μl) and DCM (188 μl) were added to theresidue and heated at 45° C. for 30 min. TFA was removed in vacuum andthe residue was purified by preparative HPLC chromatography using agradient (5 to 75%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (3.6 mg). [M+H]⁺ calcdfor C₂₆H₂₇N₉ 465.57, found 466.1.

Example 15: Synthesis ofN2-(1,3-dihydrospiro[indene-2,3′-pyrrolidin]-5-yl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(277)

Step A: Preparation of ethyl 2,3-dihydro-1H-indene-2-carboxylate (15-2).To a mixture of 15-1 (10 g, 61.6 mmol) in ethanol (300 ml) was addedH₂SO₄ (36 ml). The mixture was heated at 75° C. for 48 h. The reactionmixture was concentrated and the residue was partitioned between EtOAc(200 ml) and H₂O (200 ml). The reaction mixture was basified by 5N NaOHaq. to pH=11, then the organic layer was separated. The aqueous layerwas extracted with EtOAc (200 mL×2) and all organic layers were combinedand dried over Na₂SO₄. Concentration in vacuum gave 15-2 (10.5 g) ascolorless oil.

Step B: Preparation of ethyl 2-allyl-2,3-dihydro-1H-indene-2-carboxylate(15-4). To a solution of 15-2 (6.0 g, 31.5 mmol) in THF (120 ml) wasadded NaHMDS (38 mL, 37.8 mmol) dropwise at −78° C. The reaction mixturewas stirred at −78° C. for 1 h under N₂, then 15-3 (4.6 g, 37.8 mmol)was added at −78° C. The reaction mixture was stirred at −78° C. for 2h. The reaction mixture was diluted with sat. NH₄Cl aq (120 mL), thenextracted with EtOAC (150 ml×3). The combined organic layer wasconcentrated in vacuum and purified by silica-gel column chromatographyusing a gradient (0 to 2%) of EtOAc in PE to obtain 15-4 (6.5 g) as acolorless oil.

Step C: Preparation of ethyl2-(2-oxoethyl)-2,3-dihydro-1H-indene-2-carboxylate (15-5). A solution of15-4 (6.5 g, 28.2 mmol) in DCM (80 ml) and cooled to −78° C. O₃ wasbubbled through the solution for 15 min, at which time a light bluecolor persisted. Then TEA (5.7 g, 56.4 mmol) was added dropwise and themixture was stirred at 25° C. for 1.75 h. The reaction mixture wasdiluted with sat. NaHCO₃ aq (150 ml), then extracted with DCM (150ml×3). The combined organic layer was dried with Na₂SO₄, filtered andconcentrated in vacuum to obtain 15-5 (6.0 g) as a light yellow oil,used directly in the next step.

Step D: Preparation of 1,3-dihydrospiro[indene-2,3′-pyrrolidin]-2′-one(15-6). To a solution of 15-5 (1.0 g, 4.30 mmol) in MeOH (25 ml) andAcOH (25 ml) was added NH₄OAc (16.6 g, 215.0 mmol), the mixture wasstirred at 25° C. for 4 h. Then NaCNBH₃ (405 mg, 6.45 mmol) was addedand the mixture was stirred at 25° C. for 12 h. The reaction mixture wasdiluted with sat. NaHCO₃ aq (250 ml) and extracted with DCM (150 ml×3).The combined organic layer was dried with Na₂SO₄ and concentrated invacuum. The residue was dissolved in toluene (10 ml) and heated at 110°C. for 2 h. The reaction mixture was concentrated in vacuum and purifiedby silica-gel column chromatography using a gradient (10 to 100%) ofEtOAc in PE to obtain 15-6 (160 mg) as a white solid.

Step E: Preparation of5-nitro-1,3-dihydrospiro[indene-2,3′-pyrrolidin]-2′-one (15-7). Asolution of 15-6 (900 mg, 4.80 mmol) in HNO₃ (20 ml, 70%) was stirred at0° C. for 2 h. The reaction mixture was diluted with H₂O (30 ml) andextracted with DCM (30 ml×3). The combined organic layer wasconcentrated in vacuum and purified by silica-gel column chromatographyusing a gradient (10 to 80%) of EtOAc in PE to obtain 15-7 (700 mg) aslight yellow solid.

Step F: Preparation of 5-nitro-1,3-dihydrospiro[indene-2,3′-pyrrolidine](15-8). To a solution of 15-7 (100 mg, 0.43 mmol) in THF (4 ml) wasadded B₂H₆-Me₂S (0.8 ml, 8.60 mmol), then the mixture was heated at 45°C. for 16 h. The reaction mixture was quenched with MeOH (50 ml), thenconcentrated in vacuum and purified by preparative HPLC chromatographywith acetonitrile in water with 0.05% HCl to obtain 15-8 (50 mg) aslight yellow oil. [M+H]⁺ calcd for C₁₂H₁₄N₂O₂ 218.26, found 219.1.

Step G: Preparation of tert-butyl5-nitro-1,3-dihydrospiro[indene-2,3′-pyrrolidine]-1′-carboxylate (15-9).To a solution of 15-8 (217 mg, 0.99 mmol), DMAP (61 mg, 0.50 mmol), andTEA (1.0 g, 9.90 mmol) in THF (10 ml) was added Boc₂O (648 mg, 2.97mmol), then the reaction mixture was heated at 45° C. for 2 h. Thereaction mixture was diluted with H₂O (20 ml) and extracted with EtOAc(20 ml×3). The combined organic layer was concentrated in vacuum andpurified by silica-gel column chromatography using a gradient (0 to 20%)of EtOAc in PE to obtain 15-9 (250 mg) as light yellow oil.

Step H: Preparation of tert-butyl5-amino-1,3-dihydrospiro[indene-2,3′-pyrrolidine]-1′-carboxylate(15-10). To a solution of 15-9 (40 mg, 0.12 mmol) in MeOH (3 ml) wasadded Pd/C (10 mg, 10%), then the resulting mixture was stirred at 25°C. for 2 h under H₂ atmosphere (15 psi). The reaction mixture wasfiltered and concentrated in vacuum. The residue was purified bypreparative HPLC chromatography (acetonitrile in water with 0.225%formic acid), then the aqueous phase was adjusted to pH=8 by added sat.NaHCO₃ aq at 0° C. and the product extracted in DCM (10 ml×3) at 0° C.The combined organic layer was washed with brine (30 ml×3) at 0° C.,dried with Na₂SO₄ and concentrated in vacuum to obtain 15-9 (35 mg) asyellow solid. [M+Na]⁺ calcd for C₁₇H₂₄N₂O₂ 311.38, found 311.1.

Step I: Preparation ofN2-(1,3-dihydrospiro[indene-2,3′-pyrrolidin]-5-yl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(277). A vial of 1-8 (25 mg, 0.075 mmol), 15-10 (28 mg, 0.112 mmol),cesium carbonate (48.6 mg, 0.149 mmol), BrettPhos (8.01 mg, 0.015 mmol),and BrettPhos Pd G4 (13.73 mg, 0.015 mmol) in degassed 1,4-dioxane (1ml) was heated to 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (1 ml) and DCM (1 ml) were added to theresidue and heated at 45° C. for 30 min. TFA was removed in vacuum andthe residue was purified by preparative HPLC chromatography using agradient (5 to 65%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (16.7 mg). [M+H]⁺ calcdfor C₂₃H₂₂N₈ 410.49, found 411.1.

Example 16: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)pyrimidine-2,4-diamine(84)

Step A: Preparation of 4-(2-(4-nitro-1H-pyrazol-1-yl)ethyl)morpholine(16-3). To a stirred solution of 16-1 (0.5 g, 4.42 mmol) in DMF (5.0 ml)was added K₂CO₃ (1.83 g, 13.26 mmol) and 16-2 (1.06 g, 5.52 mmol). Thereaction mixture was stirred at 25° C. for 16 h. The reaction mixturewas diluted with ice-cold water and extracted with EtOAc (50 ml×3). Thecombined organic layer was washed with ice cold water, brine and driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto afford crude residue 16-3 (550 mg) as a viscous liquid. The crudeproduct was used directly in the next step without further purification.

Step B: Preparation of 1-(2-morpholinoethyl)-1H-pyrazol-4-amine (16-4).To a solution of 16-3 (500 mg, 2.2 mmol) in EtOAc (25 ml) was added 10%Pd/C (50 mg). The reaction mixture stirred at 25° C. for 16 h under H₂balloon pressure. The reaction mixture was filtered through a pad ofCelite and the filtrate was concentrated under reduced pressure to getcrude residue 16-4 (275 mg) as a viscous liquid. The crude product wasused directly in the next step without further purification.

Step C: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)pyrimidine-2,4-diamine(84). A vial of 1-8 (20 mg, 0.067 mmol), 16-4 (20 mg, 0.100 mmol),cesium carbonate (43.6 mg, 0.134 mmol), BrettPhos (7.19 mg, 0.013 mmol),and BrettPhos Pd G4 (12.33 mg, 0.013 mmol) in degassed 1,4-dioxane (1ml) was heated at 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (1 ml) and DCM (1 ml) were added to theresidue and stirred at 25° C. for 16 h. TFA was removed in vacuum andthe residue was purified by preparative HPLC chromatography using agradient (2 to 20%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (10 mg). [M+H]⁺ calcd forC₂₃H₂₆N₁₀O 458.53, found 459.2.

Example 17: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(1-(oxetan-3-yl)-1H-pyrazol-4-yl)phenyl)pyrimidine-2,4-diamine(538)

Step A: Preparation ofN2-(4-bromophenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(17-2). A vial of 1-8 (485 mg, 1.624 mmol), and 17-1 (559 mg, 3.25 mmol)in ethanol (8 ml) was heated at 80° C. for 16 h. The precipitate fromthe reaction mixture was collected by vacuum filtration to give thecrude residue 17-2 (620 mg), used directly in the next step withoutfurther purification. [M+H]⁺ calcd for C₂₀H₁₆BrN₇ 434.30, found 436.2.

Step B: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(1-(oxetan-3-yl)-1H-pyrazol-4-yl)phenyl)pyrimidine-2,4-diamine(538). A vial of 17-2 (25 mg, 0.058 mmol), 17-3 (21.6 mg, 0.086 mmol),potassium phosphate tribasic (36.7 mg, 0.173 mmol), palladium (II)acetate (2.58 mg, 0.012 mmol), and DPPF (5.46 mg, 0.012 mmol) indegassed 1,4-dioxane (461 μl) and H₂O (115 μl) was heated at 110° C. for16 h. The reaction mixture was concentrated in vacuum and the residuewas purified by preparative HPLC chromatography using a gradient (5 to65%) of acetonitrile in water with 0.05% trifluoroacetic acid to yield aTFA salt of the title compound (5.4 mg). [M+H]⁺ calcd for C₂₆H₂₃N₉O477.53, found 478.1.

Example 18: Synthesis ofN2-(4-(1-(2-(methylamino)ethyl)-1H-pyrazol-4-yl)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(347)

Step A: Preparation of tert-butyl(2-(4-(4-aminophenyl)-1H-pyrazol-1-yl)ethyl)(methyl)carbamate (18-2). Avial of 18-1 (96 mg, 0.274 mmol), 17-1 (39.3 mg, 0.228 mmol), potassiumcarbonate (95 mg, 0.685 mmol), and Pd(dppf)Cl₂ (33.4 mg, 0.046 mmol) indegassed 1,4-dioxane (1218 μl) and H₂O (304 μl) was heated at 110° C.for 16 h. The reaction mixture was concentrated in vacuum and theresidue was purified by preparative HPLC chromatography using a gradient(10 to 50%) of acetonitrile in water with 0.05% trifluoroacetic acid toyield a TFA salt of 18-2 (32 mg). [M+H]⁺ calcd for C₁₇H₂₄N₄O₂ 316.41,found 317.2.

Step B: Preparation ofN2-(4-(1-(2-(methylamino)ethyl)-1H-pyrazol-4-yl)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(347). A vial of 1-8 (25 mg, 0.084 mmol), 18-2 (34.4 mg, 0.109 mmol),cesium carbonate (54.5 mg, 0.167 mmol), BrettPhos (8.98 mg, 0.017 mmol),and BrettPhos Pd G4 (15.41 mg, 0.017 mmol) in degassed 1,4-dioxane (418μl) was heated at 110° C. for 16 h. The reaction mixture wasconcentrated in vacuum. TFA (226 μl) and DCM (188 μl) were added to theresidue and heated at 45° C. for 30 min. TFA was removed in vacuum andthe residue was purified by preparative HPLC chromatography using agradient (2 to 60%) of acetonitrile in water with 0.05% trifluoroaceticacid to yield a TFA salt of the title compound (26.9 mg). [M+H]⁺ calcdfor C₂₆H₂₆N₁₀ 478.56, found 479.2.

Example 19: Synthesis ofN4-(2-(3-methyl-1H-indazol-4-yl)pyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(19-6)

Step A: Preparation of bis(2-chloropyrimidin-4-yl)amine (19-2). Asuspension of 19-1 (11.5 g, 77.2 mmol), 1-7 (10.0 g, 77.2 mmol) andCs₂CO₃ (50.3 g, 154 mmol) in DMF (300 ml) was heated at 80° C. for 16 h.The reaction mixture was filtered and the filtrate was concentrated invacuum and purified by preparative HPLC chromatography using a gradient(2 to 55%) of acetonitrile in water with 0.1% trifluoroacetic acid toyield 19-2 (4.5 g) as yellow solid. [M+H]⁺ calcd for C₈H₅Cl₂N₅ 242.06,found 242.1.

Step B: Preparation ofN4-(2-chloropyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(19-3). To a suspension of 19-2 (2.0 g, 8.26 mmol) in 1,4-dioxane (80ml) was added 1-9 (982 mg, 5.51 mmol), Cs₂CO₃ (3.6 g, 11.0 mmol),Pd(OAc)₂ (123 mg, 0.551 mmol) and Xantphos (319 mg, 0.551 mmol). Thereaction mixture was degassed under vacuum and N₂ was purged for 3times. The reaction mixture was heated at 100° C. for 16 h. The reactionmixture was concentrated in vacuum and purified by silica-gel columnchromatography using a gradient (10 to 50%) of EtOAc in PE to obtain19-3 (690 mg) as yellow solid. [M+H]⁺ calcd for C₁₈H₁₈ClN₇O 383.84,found 384.0.

Step C: Preparation ofN4-(2-iodopyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(19-4). To a solution of 19-3 (690 mg, 1.80 mmol) and NaI (1.6 g, 10.8mmol) in CHCl₃ (10 ml) was added HI/H₂O (20 ml, 45%) at 0° C. and wasstirred at 25° C. for 72 h. The mixture was basified by NaOH solution (2M) to pH=8 and extracted with EtOAc (30 ml×2). The organic layer wasconcentrated in vacuum and purified by preparative HPLC chromatographyusing a gradient (10 to 35%) of acetonitrile in water with 0.225% formicacid to yield 19-4 (720 mg) as white solid. [M+H]⁺ calcd for C₁₈H₁₈IN₇O475.29, found 475.8.

Step D: Preparation ofN4-(2-(3-methyl-1H-indazol-4-yl)pyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(19-6). To a vial containing compound 19-4 (20 mg, 0.042 mmol) was added19-5 (15 mg, 0.084 mmol) followed by sodium carbonate (22.30 mg, 0.210mmol), and Pd(dppf)Cl₂ (6.16 mg, 0.0084 mmol). The resulting mixture waspurged with N₂ before degassed water (1 ml) and 1,4-dioxane (1 ml) wasadded. The vial was capped and heated at 80° C. for 16 h. The reactionwas then cooled and concentrated in vacuum. The resulting residue waspurified by preparative HPLC chromatography using a gradient (5 to 65%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (3.5 mg). [M+H]⁺ calcd for C₂₆H₂₅N₉O 479.55,found 480.1.

Example 20: Synthesis ofN4-(2-(6-(difluoromethyl)-5-fluoropyridin-2-yl)pyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(426)

Step A: Preparation of 3-fluoro-6-iodo-2-methylpyridine (20-2). To asolution of 20-1 (2.0 g, 10.5 mmol) in MeCN (10 ml) was added NaI (4.8g, 31.5 mmol) and HI aq. (2.5 ml, 47% in H₂O). The mixture was heated at135° C. for 1 h under microwave. The reaction was poured into cold sat.NaHCO₃ (300 ml). The resulting mixture was extracted with EtOAc (150ml×3). The organic layers were washed with sat. Na₂SO₃ (200 ml×3) andconcentrated in vacuum to dryness to give crude residue 20-2 (2.0 g) aswhite solid, used directly in the next step. [M+H]⁺ calcd for C₆H₅FIN237.02, found 237.9.

Step B: Preparation of 3-fluoro-6-iodo-2-methylpyridine 1-oxide (20-3).To a solution of 20-2 (10 g, 42.2 mmol) in toluene (200 ml) was addedmCPBA (17.1 g, 84.4 mmol, 85% purity) in portions. The mixture wasstirred at 25° C. for 2 h. The reaction mixture was poured into icesaturated NaHCO₃ aq. (500 ml). The resulting mixture was extracted withEtOAc (500 ml×5). The combined organic layer was washed with Na₂SO₃ aq.(1000 ml×3), concentrated to dryness and purified by column (PE/EA=20/1to 1/1) to give 20-3 (4.67 g) as yellow solid. [M+H]⁺ calcd for C₆H₅FINO253.01, found 253.9.

Step C: Preparation of (3-fluoro-6-iodopyridin-2-yl)methanol (20-4).20-3 (10.5 g, 41.49 mmol) was added into TFAA (100 ml) in batches. Thereaction mixture was stirred at 35° C. for 16 h. The reaction mixturewas poured into ice NaHCO₃ aq. (500 ml). The resulting mixture wasextracted with EtOAc (200 ml×3). The combined organic layer wasconcentrated to dryness to give a crude product. This crude product wasdissolved in DCM (80 ml) and H₂O (80 ml). Then K₂CO₃ (22.9 g, 165.97mmol) was added. The reaction mixture was heated at 40° C. for 16 h. Thereaction mixture was poured into ice water (300 ml). The resultingmixture was extracted with EtOAc (200 ml×3). The combined organic layerwas concentrated to dryness and purified by column (PE/EA=100/1 to 10/1)to obtain 20-4 (7.3 g) as light yellow solid. [M+H]⁺ calcd for C₆H₅FINO253.01, found 253.9.

Step D: Preparation of 3-fluoro-6-iodopicolinaldehyde (20-5). To asolution of 20-4 (7.3 g, 28.9 mmol) in DCM (160 ml) was added DMP (24.5g, 57.8 mmol). The mixture was stirred at 25° C. for 2 h. The reactionwas poured into cold sat. NaHCO₃ (500 ml). The mixture was extractedwith EtOAc (200 ml×5). The organic layer was concentrated in vacuum todryness and purified by column (PE/EA=100/1 to 10/1) to obtain 20-5 (6.5g) as light yellow solid.

Step E: Preparation of 2-(difluoromethyl)-3-fluoro-6-iodopyridine(20-6). To a solution of 20-5 (6.5 g, 25.9 mmol) in DCM (100 ml) wasdropwise added DAST (7.5 g, 46.6 mmol) at −20° C. The reaction mixturewas warmed slowly and stirred at 20° C. for another 2 h. The reactionmixture was neutralized with sat. NaHCO₃ aq. (500 ml), then theresulting mixture was extracted with EtOAc (150 ml×3). The combinedorganic layers were concentrated to dryness and purified by silica-gelcolumn chromatography (PE/EA=100/1 to 10/1) to obtain 20-6 (6.5 g) aslight pink solid. [M+H]⁺ calcd for C₆H₃F₃IN 273.00, found 273.9.

Step F: Preparation of (6-(difluoromethyl)-5-fluoropyridin-2-yl)boronicacid (20-8). To a solution of 20-6 (1.8 g, 6.59 mmol) and B(OPr-i)₃ (2.5g, 13.4 mmol) in THF (70 ml) was dropwise added i-PrMgCl (16.5 mL, 32.9mmol, 2.0 M/L) at 0° C. The reaction mixture was warmed and stirred at25° C. for 16 h. The reaction mixture was quenched with H₂O (100 ml).The resulting reaction mixture was extracted with MTBE (50 ml×3). Theaqueous phase was lyophilized to give a crude product. This crudeproduct was triturated with EtOAc (100 ml×3) for 2 h at 25° C. Themother liquor was lyophilized again, and then added to H₂O (50 ml) andextracted with MTBE (50 ml×3) and PE (50 ml×3). The aqueous phase waslyophilized again to give crude residue 20-8 (720 mg) as white solid,used directly in the next step. [M+H]⁺ calcd for C₆H₅BF₃NO₂ 190.92,found 191.3.

Step G: Preparation ofN4-(2-(6-(difluoromethyl)-5-fluoropyridin-2-yl)pyrimidin-4-yl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine(426). To a vial containing 19-6 (20 mg, 0.042 mmol) was added 20-8 (16mg, 0.084 mmol) followed by sodium carbonate (22.30 mg, 0.210 mmol) andPd(dppf)Cl₂ (6.16 mg, 0.0084 mmol). The resulting mixture was purgedwith N₂ before degassed water (1 ml) and 1,4-dioxane (1 ml) was added.The vial was capped and heated at 80° C. for 16 h. The reaction was thencooled and concentrated in vacuum. The resulting residue was purified bypreparative HPLC chromatography using a gradient (15 to 75%) ofacetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (4.7 mg). [M+H]⁺ calcd for C₂₄H₂₁F₃N₈O494.48, found 495.0.

Example 21: Synthesis ofN4-(2-(5-chloro-2,4-difluorophenyl)pyrimidin-4-yl)-N2-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4-diamine(556)

Step A: Preparation of tert-butyl4-(4-((4-((2-chloropyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazine-1-carboxylate(21-1). To a suspension of 19-2 (2.6 g, 10.8 mmol) in 1,4-dioxane (72ml) was added 2-1 (2.0 g, 7.21 mmol), Cs₂CO₃ (4.7 g, 14.2 mmol),Pd(OAc)₂ (162 mg, 0.721 mmol) and Xantphos (417 mg, 0.721 mmol). Thereaction mixture was degassed under vacuum and N₂ was purged 3 times.The mixture was heated at 100° C. for 16 h. The reaction mixture wasconcentrated in vacuum and purified by silica gel column chromatographyusing a gradient (20 to 50%) of EtOAc in PE to obtain 21-1 (730 mg) asyellow solid. [M+H]⁺ calcd for C₂₃H₂₇ClN₈O₂ 482.97, found 483.4.

Step B: Preparation ofN4-(2-chloropyrimidin-4-yl)-N2-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4-diamine(21-2). To a solution of 21-1 (730 mg, 1.51 mmol) in DCM (8 ml) wasadded TFA (1.0 g, 15.1 mmol) and the resulting mixture was stirred at25° C. for 3 h. The reaction mixture was concentrated in vacuum and theresidue was basified by saturated NaHCO₃ solution to pH=8. The reactionmixture was filtered and the filtrate was dried in vacuum to obtain 21-2(520 mg) as white solid, used directly in the next step. [M+H]⁺ calcdfor C₁₈H₁₉ClN₈ 382.86, found 383.2.

Step C: Preparation ofN4-(2-iodopyrimidin-4-yl)-N2-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4-diamine(21-3). To a solution of 21-2 (520 mg, 1.36 mmol) and NaI (1.2 g, 8.15mmol) in CHCl₃ (10 ml) was added HI/H₂O (20 ml, 45%) at 0° C. and theresulting mixture was stirred at 25° C. for 7 d. The reaction mixturewas basified by sat. NaHCO₃ solution to pH=8 at 0° C., washed withsaturated Na₂SO₃ solution (20 ml) and extracted with EtOAc (30 ml×3).The organic layer was concentrated in vacuum and purified by preparativeHPLC chromatography using a gradient (10 to 40%) of acetonitrile inwater with 0.05% HCl to obtain 21-3 (120 mg) as yellow solid. [M+H]⁺calcd for C₁₈H₁₉1N₈ 474.31, found 475.83.

Step D: Preparation ofN4-(2-(5-chloro-2,4-difluorophenyl)pyrimidin-4-yl)-N2-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4-diamine(556). To a vial containing compound 21-3 (10 mg, 0.021 mmol) was added21-4 (8.08 mg, 0.042 mmol) followed by sodium carbonate (11.17 mg, 0.105mmol) and Pd(dppf)Cl₂ (3.09 mg, 0.0042 mmol). The resulting mixture waspurged with N₂ before degassed water (290 μl) and 1,4-dioxane (290 μl)was added. The vial was capped and heated at 110° C. for 1 h. Thereaction was then cooled and concentrated in vacuum. The resultingresidue was purified by preparative HPLC chromatography using a gradient(15 to 75%) of acetonitrile in water with 0.05% trifluoroacetic acid toyield a TFA salt of the title compound (5.7 mg). [M+H]⁺ calcd forC₂₄H₂₁ClF₂N₈ 494.94, found 495.1.

Example 22: Synthesis of4-methyl-3-(4-((2-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)pyrimidin-2-yl)phenol(255)

Step A: Preparation of tert-butylmethyl(1-(4-nitrophenyl)piperidin-4-yl)carbamate (22-2). To a solutionof 5-1 (4.0 g, 18.7 mmol) in DMF (60 ml) was added 22-1 (2.6 g, 18.7mmol) and K₂CO₃ (5.2 g, 37.3 mmol). The reaction mixture was stirred at25° C. for 15 h. The reaction was filtrated and the cake was washed withEtOAc (80 ml×2). The filtrate was poured into 250 ml of H₂O. The mixturewas extracted with EtOAc (150 ml×3). The organic layer was washed withsat. NaCl (100 ml×3), dried over with Na₂SO₄ and concentrated in vacuumto dryness to give crude product. The crude product was purified bycolumn (PE:EA=50:1 to 3:1) to give 22-2 (5.8 g) as yellow solid.

Step B: Preparation of tert-butyl(1-(4-aminophenyl)piperidin-4-yl)(methyl)carbamate (22-3). To a solutionof 22-2 (5.7 g, 16.99 mmol) in MeOH (80 ml) was added Pd/C (500 mg,10%). The reaction mixture was stirred at 25° C. for 15 h under H₂balloon. The reaction mixture was filtrated through a pad of Celite andthe solvent was removed in vacuum to dryness to give crude residue 22-3(4.5 g) as light pink solid, used directly in the next step.

Step C: Preparation of tert-butyl(1-(4-((4-((2-chloropyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)carbamate(22-4). A suspension of 19-3 (95 mg, 0.393 mmol), 22-3 (100 mg, 0.327mmol), Pd(OAc)₂ (7 mg, 0.033 mmol), Xantphos (19 mg, 0.033 mmol) andCs₂CO₃ (213 mg, 0.655 mmol) in 1,4-dioxane (9 ml) was heated at 120° C.under microwave for 3 h under N₂. The reaction mixture was filtrated andthe filtrate was purified by preparative HPLC chromatography using agradient (15 to 45%) of acetonitrile in water with 0.1% trifluoroaceticacid to give 22-4 (400 mg) as brown solid. [M+H]⁺ calcd for C₂₅H₃₁ClN₈O₂511.03, found 511.4.

Step D: Preparation ofN4-(2-chloropyrimidin-4-yl)-N2-(4-(4-(methylamino)piperidin-1-yl)phenyl)pyrimidine-2,4-diamine(22-5). A suspension of 22-4 (400 mg, 0.783 mmol) in HCl/EA (25 ml, 4M)was stirred at 0° C. for 1 h. The solvent was removed in vacuum to givecrude residue 22-5 (500 mg) as pink solid, used directly in the nextstep. [M+H]⁺ calcd for C₂₀H₂₃ClN₈ 410.91, found 410.3.

Step E: Preparation ofN4-(2-iodopyrimidin-4-yl)-N2-(4-(4-(methylamino)piperidin-1-yl)phenyl)pyrimidine-2,4-diamine(22-6). To a suspension of 22-5 (400 mg, 0.973 mmol) in CHCl₃ (5 ml) wasadded HI/H₂O (30 ml, 45% in H₂O). The reaction mixture was stirred at25° C. for 11 d. The reaction was poured into 100 ml of ice water at 0°C. 15 g of Na₂SO₃ was added to the solution. pH of the solution wasadjusted to 8 with sat. NaHCO₃. The solution was extracted with EtOAc(50 ml×6). The organic layer was dried with Na₂SO₄ and concentrated invacuum to dryness to give crude residue. The residue was purified bypreparative HPLC chromatography using a gradient (7 to 37%) ofacetonitrile in water with 0.05% HCl to give the HCl salt of 22-6 (180mg) as yellow solid. [M+H]⁺ calcd for C₂₀H₂₃IN₈ 502.36, found 503.2.

Step F: Preparation of4-methyl-3-(4-((2-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)pyrimidin-2-yl)phenol(255). To a vial containing compound 22-6 (15 mg, 0.026 mmol) was added22-7 (12 mg, 0.079 mmol) followed by sodium carbonate (13.91 mg, 0.131mmol), and Pd(dppf)Cl₂ (3.85 mg, 0.0052 mmol). The resulting mixture waspurged with N₂ before degassed water (0.5 ml) and 1,4-dioxane (1 ml) wasadded. The vial was capped and heated at 100° C. for 16 h. The reactionwas then cooled and concentrated in vacuum. The resulting residue waspurified by preparative HPLC chromatography using a gradient (5 to 65%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (14.5 mg). [M+H]⁺ calcd for C₃₁H₃₇N₉O 551.70,found 552.2.

Example 23: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(piperazin-1-ylmethyl)thiazol-2-yl)pyrimidine-2,4-diamine(287)

Step A: Preparation of tert-butyl4-((2-aminothiazol-4-yl)methyl)piperazine-1-carboxylate (23-3). To asolution of 23-1 (150 mg, 0.807 mmol), and 23-2 (100 mg, 0.673 mmol) inDMF (2.7 ml) was added DIEA (0.59 ml, 3.36 mmol). The reaction mixturewas heated at 110° C. for 1 h. The reaction mixture was concentrated invacuum and was purified by silica-gel column chromatography using agradient (0 to 15%) of MeOH in DCM to obtain 23-3 (120 mg) as yellowoil. [M+H]⁺ calcd for C₁₃H₂₂N₄O₂S 298.41, found 299.4.

Step B: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(4-(piperazin-1-ylmethyl)thiazol-2-yl)pyrimidine-2,4-diamine(287). A vial of 1-8 (100 mg, 0.335 mmol), 23-3 (120 mg, 0.402 mmol),cesium carbonate (218 mg, 0.669 mmol), Xantphos (38.7 mg, 0.067 mmol),and Pd(OAc)₂ (15.03 mg, 0.067 mmol) in degassed 1,4-dioxane (3.3 ml) washeated at 110° C. for 16 h. The reaction mixture was concentrated invacuum. TFA (903 μl) and DCM (754 μl) were added to the residue andstirred at 45° C. for 30 min TFA was removed in vacuum and the residuewas purified by preparative HPLC chromatography using a gradient (5 to60%) of acetonitrile in water with 0.05% trifluoroacetic acid to yield aTFA salt of the title compound (124 mg). [M+H]⁺ calcd for C₂₂H₂₄N₁₀S460.56, found 461.2.

Example 24: Synthesis ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(5-(piperazin-1-ylmethyl)thiophen-3-yl)pyrimidine-2,4-diamine(94)

Step A: Preparation of methyl4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-2-carboxylate(24-2). To a solution of 1-8 (800 mg, 5.09 mmol) in 1,4-dioxane (10 nil)was added 24-1 (1.21 g, 4.07 mmol) under N₂ atmosphere. Then Cs₂CO₃(3.30 g, 10.18 mmol) was added. The reaction mixture was purged with N₂for 5 min, followed by addition of Pd₂(dba)₃ (233 mg, 0.25 mmol) andBINAP (317 mg, 0.05 mmol), then heated at 110° C. for 6 h in a sealedvial. The reaction mixture was diluted with EtOAc, filtered through aCelite bed, and concentrated to obtain the crude residue. The residuewas purified by silica-gel column chromatography using a gradient (0 to3%) of MeOH in DCM to obtain 24-2 (820 mg) as yellow solid. [M+H]⁺ calcdfor C₂₀H₁₇N₇O₂S 419.46, found 420.12.

Step B: Preparation of(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophen-2-yl)methanol(24-3). To a solution of 24-2 (800 mg, 1.90 mmol) in dry THF (10 ml) wasadded LAH (1.90 ml, 3.81 mmol) under N₂ atmosphere at 0° C. The reactionstirred at 25° C. for 2 h. The reaction mixture was cooled to −30° C.and quenched with saturated ammonium chloride solution, diluted withEtOAc and filtered through Celite bed, then concentrated to get cruderesidue. The residue was purified by silica-gel column chromatographyusing a gradient (0 to 8%) of MeOH in DCM to obtain 24-3 (300 mg) asoff-white solid. [M+H]⁺ calcd for C₁₉H₁₇N₇OS 391.45, found 392.15.

Step C: Preparation of4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-2-carbaldehyde(24-4). To a solution of 24-3 (250 mg, 0.63 mmol) in DCM (8 ml) wasadded MnO₂ (1.39 g, 15.98 mmol) at 25° C. The resulting suspension wasstirred at 25° C. for 16 h. The reaction mixture was diluted with DCM,filtered through Celite bed, and concentrated to afford the cruderesidue which was purified by washing with diethyl ether and ACN toobtain 24-4 (180 mg) as yellow solid. [M+H]⁺ calcd for C₁₉H₁₅N₇OS389.44, found 390.27.

Step D: Preparation ofN4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)-N2-(5-(piperazin-1-ylmethyl)thiophen-3-yl)pyrimidine-2,4-diamine(94). To a stirred solution 24-4 (75 mg, 0.193 mmol) in DMA (800 μl) wasadded 23-1 (30 mg, 0.161 mmol) followed by the addition of STAB (102 mg,0.483 mmol). The reaction mixture was stirred at 25° C. for 16 h. Thereaction mixture was concentrated in vacuum. DCM (400 μl) and TFA (400μl) were added to the reaction mixture and the resulting mixture heatedat 45° C. for 30 min. TFA was removed in vacuum. The residue waspurified by preparative HPLC chromatography using a gradient (2 to 15%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (29.6 mg). [M+H]⁺ calcd for C₂₃H₂₅N₉S 459.58,found 460.2.

Example 25: Synthesis ofN-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)-1-(piperidin-3-yl)methanesulfonamide(106)

To a vial of 17-2 (20 mg, 0.054 mmol) and 25-1 (24 mg, 0.081 mmol) inDMF (1 ml) was added DIEA (0.028 ml, 0.162 mmol) and the resultingmixture was stirred at 25° C. for 16 h. TFA (1 ml) was added to thereaction mixture and heated at 50° C. for 1 h. The reaction mixture wasconcentrated in vacuum and the residue was purified by preparative HPLCchromatography using a gradient (2 to 60%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(10.3 mg). [M+H]⁺ calcd for C₂₆H₂₉N₉O₂S 531.64, found 532.1.

Example 26: Synthesis ofN-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazine-1-carboxamide(228)

To a solution of 17-2 (25 mg, 0.067 mmol) in DCM (1 ml) was added CDI(12.04 mg, 0.074 mmol) and the resulting mixture stirred at 25° C. for16 h. 23-1 (13.83 mg, 0.074 mmol) was added to the reaction mixture andstirred at 25° C. for 16 h. The reaction mixture was concentrated invacuum. TFA (1 ml) and DCM (1 ml) were added to the residue and heatedat 45° C. for 30 min. TFA was removed in vacuum and the residue waspurified by preparative HPLC chromatography using a gradient (5 to 65%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (11.8 mg). [M+H]⁺ calcd for C₂₅H₂₆N₁₀O482.55, found 483.1.

Example 27: Synthesis ofN-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperidine-4-carboxamide(14)

To a solution of 17-2 (12 mg, 0.032 mmol) and 27-1 (14.86 mg, 0.065mmol) in DMF (1 ml) was added HATU (18.48 mg, 0.049 mmol) and DIEA(0.017 ml, 0.097 mmol) and the resulting mixture stirred at 25° C. for16 h. The reaction mixture was concentrated in vacuum. TFA (1 ml) andDCM (1 ml) were added to the residue and heated at 45° C. for 30 min.TFA was removed in vacuum and the residue was purified by preparativeHPLC chromatography using a gradient (5 to 65%) of acetonitrile in waterwith 0.05% trifluoroacetic acid to yield a TFA salt of the titlecompound (19.7 mg). [M+H]⁺ calcd for C₂₆H₂₇N₉O 481.56, found 482.1.

Example 28: Synthesis of3-(hydroxymethyl)-N-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)azetidine-3-carboxamide(209)

To a solution of 17-2 (20 mg, 0.054 mmol) and 28-1 (18.73 mg, 0.081mmol) in DMF (1 ml) was added HATU (30.8 mg, 0.081 mmol) and DIEA (0.028ml, 0.162 mmol) and the resulting mixture was stirred at 25° C. for 1 h.The reaction mixture was concentrated in vacuum. TFA (229 μl) and DCM(191 μl) were added to the residue and heated at 45° C. for 30 min. TFAwas removed in vacuum and the residue was purified by preparative HPLCchromatography using a gradient (2 to 60%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(35.1 mg). [M+H]⁺ calcd for C₂₅H₂₅N₉O₂ 483.54, found 484.0.

Example 29: Synthesis ofN-(3-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperidine-3-carboxamide(443)

Step A: Preparation ofN2-(3-aminophenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(29-2). To a mixture of 1-8 (400 mg, 1.34 mmol) and 29-1 (144 mg, 1.34mmol) in IPA (10 ml) was added HCl (0.5 ml). The reaction mixture washeated at 80° C. for 16 h. The reaction mixture was filtered. Thefiltrate was concentrated in vacuum and purified by silica-gel columnchromatography using a gradient (0 to 5%) of MeOH in DCM to give 29-2(280 mg) as yellow solid. [M+H]⁺ calcd for C₂₀H₁₈N₈ 370.42, found 371.2.

Step B: Preparation ofN-(3-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperidine-3-carboxamide(443). To a solution of 29-2 (20 mg, 0.054 mmol) and 29-3 (18.73 mg,0.081 mmol) in DMF (1 ml) was added HATU (30.8 mg, 0.081 mmol) and DIEA(0.028 ml, 0.162 mmol) and the resulting mixture was stirred at 25° C.for 1 h. The reaction mixture was concentrated in vacuum. TFA (229 μl)and DCM (191 μl) were added to the residue and heated at 45° C. for 30min. TFA was removed in vacuum and the residue was purified bypreparative HPLC chromatography using a gradient (2 to 60%) ofacetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (35.1 mg). [M+H]⁺ calcd for C₂₅H₂₅N₉O₂483.54, found 484.0.

Example 30: Synthesis ofN-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophen-2-yl)piperidine-4-carboxamide(452)

Step A: Preparation of4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-2-carboxylicacid (30-1). A mixture of 24-2 (1.3 g, 3.10 mmol) and NaOH (1.2 g, 31.0mmol) in MeOH (30 ml) and H₂O (15 ml) was heated at 50° C. for 2 h. Thereaction mixture was acidified with AcOH to pH=7 and filtered. The cakewas dried in vacuum to afford 30-1 (1.2 g) as a yellow solid. [M+H]⁺calcd for C₁₉H₁₅N₇O₂S 405.44, found 406.0.

Step B: Preparation ofN2-(5-isocyanatothiophen-3-yl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(30-3). To a solution of 30-1 (1.0 g, 2.46 mmol) and TEA (1.5 g, 14.8mmol) in THF (20 ml) was added 30-2 (2.1 g, 22.3 mmol) at 0° C. Thereaction mixture was stirred at 0° C. for 6 h. Then NaN₃ (1.0 g, 15.4mmol) in H₂O (5.0 ml) was added at 0° C. The reaction mixture wasstirred at 20° C. for 12 h. The reaction mixture was concentrated invacuum. The residue was washed with H₂O (50 ml×2) and the cake was driedin vacuum to afford 30-3 (900 mg) as a white solid. [M+H]⁺ calcd forC₁₉H₁₄N₈OS 402.44, found 403.0.

Step C: Preparation ofN2-(5-aminothiophen-3-yl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(30-4). A solution of 30-3 (800 mg, 1.19 mmol) in HCl (10 ml, 6.0 M) wasstirred at 50° C. for 12 h. The reaction mixture was concentrated invacuum. The residue was purified by preparative HPLC chromatographyusing a gradient (0 to 32%) of acetonitrile in water with 0.05% HCl toobtain 30-4 (120 mg) as a white solid. [M+H]⁺ calcd for C₁₈H₁₆N₈S376.44, found 377.1.

Step D: Preparation ofN-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophen-2-yl)piperidine-4-carboxamide(452). A solution of 30-4 (20 mg, 0.053 mmol), 27-1 (18.27 mg, 0.080mmol), HATU (30.3 mg, 0.080 mmol), and DIEA (28.0 μl, 0.159 mmol) in DMF(1 ml) was stirred at 25° C. for 1 h. The reaction mixture wasconcentrated in vacuum. DCM (200 μl) and TFA (225 μl) were added to thereaction mixture and the mixture was heated at 45° C. for 30 min. TFAwas removed in vacuum and the crude mixture purified by preparative HPLCchromatography using a gradient (2 to 60%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(26.9 mg). [M+H]⁺ calcd for C₂₄H₂₅N₉OS 487.59, found 488.0.

Example 31: Synthesis of (R)-sec-butyl2-(5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)-2-(piperazin-1-yl)phenyl)acetate(13)

Step A: Preparation of methyl 2-(2-fluoro-5-nitrophenyl)acetate (31-2).To a stirred solution of 31-1 (10 g, 50.25 mmol) in MeOH (70 ml) wasadded H₂SO₄ (5.3 ml, 100 mmol) at 0° C. under N₂ atmosphere. Thereaction mixture was refluxed for 16 h. Solvent was removed underreduced pressure and was carefully quenched using bicarbonate solutionand extracted with EtOAc. The organic layer was dried over anhydroussodium sulfate and concentrated under reduced pressure to obtain thecrude product 31-2 (10 g). [M+Na]⁺ calcd for C₉H₈FNO₄ 236.15, found236.31.

Step B: Preparation of tert-butyl4-(2-(2-methoxy-2-oxoethyl)-4-nitrophenyl)piperazine-1-carboxylate(31-3). To a stirred solution of 31-2 (5.0 g, 23.47 mmol) in DMA (70 ml)was added DIPEA (12.61 ml, 70.41 mmol) followed by 23-1 (4.3 g, 23.47mmol). The reaction mixture was heated at 100° C. for 16 h. The reactionmixture was then diluted with water and extracted with EtOAc (100 ml×3).The combined organic layer was again washed with ice cold water followedby brine and dried over anhydrous Na₂SO₄, filtered and concentratedunder reduced pressure. Crude residue obtained was purified bysilica-gel column chromatography using a gradient (20 to 25%) of acetonein hexanes to obtain 31-3 (4.05 g) as a white solid. [M+H]⁺ calcd forC₁₈H₂₅N₃O₆ 379.41, found 380.34.

Step C: Preparation of2-(2-(4-(tert-butoxycarbonyl)piperazin-1-yl)-5-nitrophenyl)acetic acid(31-4). To a stirred solution of 31-3 (1.0 g, 2.63 mmol) in THF (5 ml)and H₂O (5 ml) was added LiOH·H₂O (540 mg, 13.19 mmol). The reactionmixture was stirred at 25° C. for 16 h, then diluted with water andextracted with EtOAc at pH ˜3. The organic layer was washed with brine,dried over sodium sulphate and concentrated under reduced pressure togive the crude residue. The residue was purified by precipitating andtriturating with ACN, diethyl ether and hexane to obtain 31-4 (600 mg)as a yellow solid. [M+H]⁺ calcd for C₁₇H₂₃N₃O₆ 365.39, found 366.16.

Step D: Preparation of tert-butyl(R)-4-(2-(2-(sec-butoxy)-2-oxoethyl)-4-nitrophenyl)piperazine-1-carboxylate(31-6). To a stirred solution of 31-4 (550 mg, 1.50 mmol) in DCM (20 ml)was added 31-5 (0.37 ml, 3.01 mmol) and DMAP (18 mg, 0.15 mmol) followedby addition of EDCHCl (573 mg, 3.01 mmol) at 0° C. The reaction mixturewas allowed to stir at 25° C. for 16 h, then diluted with water andextracted three times using EtOAc. The combined organic layers werewashed with water and brine, dried over anhydrous sodium sulfate andconcentrated under vacuum to afford crude compound which was purified bysilica-gel column chromatography using a gradient (10 to 15%) of EtOAcin Hexanes to obtain 31-6 (400 mg). [M+H]⁺ calcd for C₂₁H₃₁N₃O₆ 421.49,found 422.10.

Step E: Preparation of tert-butyl(R)-4-(4-amino-2-(2-(sec-butoxy)-2-oxoethyl)phenyl)piperazine-1-carboxylate(31-7). To a stirred solution of 31-6 (400 mg, 0.95 mmol) in IPA (10 ml)and THF (4 ml) was added 10% Pd/C (400 mg) and subjected tohydrogenation under H₂ balloon pressure. The reaction mixture wasallowed to stir at 25° C. for 1 h, then filtered through Celite bed andwashed with MeOH. Filtrate was concentrated under vacuum to get cruderesidue 31-7 (350 mg), used in next step without any purification.[M+H]⁺ calcd for C₂₁H₃₃N₃O₄ 391.51, found 392.13.

Step F: Preparation of tert-butyl(R)-4-(2-(2-(sec-butoxy)-2-oxoethyl)-4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazine-1-carboxylate(31-8). To a solution of 31-7 (344 mg, 0.88 mmol) in 1,4-dioxane (10 ml)was added 1-8 (175 mg, 0.587 mmol) and Cs₂CO₃ (381 mg, 1.17 mmol) underN₂ atmosphere. The reaction mixture was degassed with N₂ for 10 min,followed by addition of Brettphos (63 mg, 0.117 mmol) and Brettphos PdG4 (54 mg, 0.058 mmol), then heated at 110° C. for 16 h. The reactionmixture was diluted with EtOAc and filtered through Celite bed. Filtratewas diluted with cold water and extracted with ethyl acetate (25 mL×2).The organic layer was dried over dry Na₂SO₄, filtered and concentratedto get crude residue. The residue was purified by silica-gel columnchromatography using a gradient (0 to 3%) of MeOH in DCM to obtain 31-8(200 mg). [M+H]⁺ calcd for C₃₅H₄₃N₉O₄ 653.79, found 654.17.

Step G: Preparation of (R)-sec-butyl2-(5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)-2-(piperazin-1-yl)phenyl)acetate(13). To a stirred solution of 31-8 (200 mg, 0.306 mmol) in DCM (5 ml)was added TFA (1 ml) at 0° C. The reaction mixture was stirred at 25°C., then concentrated under reduced pressure and further triturated withdiethyl ether to get the title compound (205 mg) as an off white solid.[M+H]⁺ calcd for C₃₀H₃₅N₉O₂ 553.67, found 554.32.

Example 32: Synthesis of (R)-1-methylpyrrolidin-3-yl1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-4-carboxylate(414)

Step A: Preparation of methyl4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoate(32-2). To a stirred solution of 1-8 (3.5 g, 11.74 mmol) and 32-1 (2.3g, 15.27 mmol) in n-butanol (35 ml) was added PTSA (1.11 g, 5.87 mmol)and the resulting mixture heated at 110° C. for 3 h. The reactionmixture was concentrated to dryness and triturated using diethyl etherto get the 32-2 (4.1 g) as PTSA salt. [M+H]⁺ calcd for C₂₂H₁₉N₇O₂413.44, found 414.15.

Step B: Preparation of(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)methanol(32-3). To a stirred solution 32-2 (4.1 g, 9.92 mmol) in THF (50 ml) wasadded LAH (15.0 ml, 29.71 mmol) dropwise at 0° C. The reaction mixturewas allowed to stir at 0° C. for 2 h, then quenched with water:THF (6:18ml) under cooling followed by the addition of 15% NaOH (15 ml). It wasthen filtered through Celite bed and the organic layer separated andconcentrated to get crude residue. The residue was purified bysilica-gel column chromatography to obtain 32-3 (3.0 g). [M+H]⁺ calcdfor C₂₁H₁₉N₇O 385.43, found 386.14.

Step C: Preparation of4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzaldehyde(32-4). To a stirred solution 32-3 (3.0 g, 7.8 mmol) in DMA (30 ml) wasadded MnO₂ (20.4 g, 234 mmol) at 25° C. The reaction mixture was allowedto stir at 25° C. for 6 h, then filtered and the filtrate concentratedto get crude residue. The residue obtained was triturated with ACN toobtain 32-4 (2.2 g), used in next step without further purification.[M+H]⁺ calcd for C₂₁H₁₇N₇O 383.42, found 384.16.

Step D: Preparation of tert-butyl1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-4-carboxylate(32-6). To a stirred solution of 32-4 (2.1 g, 5.48 mmol) in DMA:DCE(10:20 ml) was added 32-5 (1.52 g, 8.22 mmol) followed by the additionof STAB (3.48 g, 16.44 mmol). The reaction mixture was stirred at 25° C.for 16 h. The reaction mixture was diluted with water and extracted with10% MeOH:DCM to obtained the crude residue. The residue was purified bysilica-gel column chromatography using a gradient (0 to 5%) of MeOH inDCM to obtain 32-6 (1.4 g). [M+H]⁺ calcd for C₃₁H₃₆N₈O₂ 552.68, found553.33.

Step E: Preparation of1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-4-carboxylicacid (32-7). To a stirred solution 32-6 (1.4 g, 2.53 mmol) in DCM (20ml) was added TFA (14 ml) dropwise at 0° C. The reaction mixture wasstirred at 0° C. for 6 h. The reaction mixture was concentrated andtriturated with diethyl ether and ACN to obtain 32-7 (1.2 g). [M+H]⁺calcd for C₂₇H₂₈N₈O₂ 496.58, found 497.48.

Step F: Preparation of (R)-1-methylpyrrolidin-3-yl1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-4-carboxylate(414). To the solution of, 32-7 (300 mg, 0.604 mmol) in DMF (5.0 ml) wasadded 32-8 (170 mg, 0.906 mmol), DCC (373 mg, 1.812 mmol) followed byaddition of DMAP (37 mg, 0.302 mmol) and stirred at 25° C. for 4 h. Thereaction mixture was poured onto ice water and extracted using 10%MeOH:DCM. Combined organic layers were washed with brine, dried oversodium sulfate and concentrated to get the crude residue. The residuewas purified by preparative HPLC chromatography using a gradient (5 to25%) of acetonitrile in water with 0.1% trifluoroacetic acid to yield aTFA salt of the title compound (180 mg). [M+H]⁺ calcd for C₃₂H₃₇N₉O₂579.71, found 580.30.

Example 33: Synthesis of azetidin-3-ylmethyl(R)-1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-3-carboxylate(400)

Step A: Preparation ofN2-(4-(chloromethyl)phenyl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(33-1). To a solution of 32-2 (425 mg, 1.103 mmol) in DCM (31.5 ml) wasadded SOCl₂ (177 μl, 2.426 mmol) and the resulting mixture heated at 60°C. for 1 h. The reaction mixture was concentrated in vacuum to obtain33-1 (445 mg) as a yellow solid, used directly in the next step.

Step B: Preparation of tert-butyl(R)-1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-3-carboxylate(33-3). To a solution of 33-1 (445 mg, 1.1 mmol) in DCM (18 ml) wasadded 33-2 (488 mg, 2.2 mmol) and DIEA (0.961 ml, 5.0 mmol) and theresulting mixture heated at 55° C. for 16 h. The reaction mixture wasconcentrated in vacuum to give a crude residue. The residue was purifiedby silica-gel column chromatography using a gradient (0 to 15%) of MeOHin DCM to produce 33-3 (608 mg). [M+H]⁺ calcd for C₃₁H₃₆N₈O₂ 552.68,found 553.1.

Step C: Preparation of(R)-1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-3-carboxylicacid (33-4). To a solution of 33-3 (608 mg, 1.1 mmol) in DCM (3.5 ml)was added TFA (4.2 ml) at 0° C. and the reaction mixture was stirred at25° C. for 16 h. TFA was removed in vacuum. The residue was purified bypreparative HPLC chromatography using a gradient (5 to 30%) ofacetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of 33-4 (440 mg). [M+H]⁺ calcd for C₂₇H₂₈N₈O₂ 496.58, found 497.1.

Step D: Preparation of azetidin-3-ylmethyl(R)-1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)piperidine-3-carboxylate(400). A solution of 33-4 (75 mg, 0.104 mmol), 33-5 (38.8 mg, 0.207mmol), EDCI (39.7 mg, 0.207 mmol), and DMAP (1.27 mg, 10.4 μmol) in DCM(0.5 ml) was stirred at 25° C. for 16 h. TFA (279 μl) was added to thereaction mixture and was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (2 to 60%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(62.1 mg). [M+H]⁺ calcd for C₃₁H₃₅N₉O₂ 565.68, found 566.1.

Example 34: Synthesis of azetidin-3-ylmethyl1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)azetidine-3-carboxylate(88)

Step A: Preparation of1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)azetidine-3-carboxylicacid (34-2). To a solution of 33-1 (262 mg, 0.65 mmol) in DCM (10 ml)was added 34-1 (204 mg, 1.30 mmol) and DIEA (0.567 ml, 3.25 mmol) andthe resulting mixture heated at 55° C. for 16 h. The reaction mixturewas concentrated in vacuum to give a crude residue. To the residue wasdirectly added DCM (2 ml) and TFA (2 ml) and was heated at 55° C. for 1h. The reaction mixture was concentrated in vacuum and was purified bypreparative HPLC chromatography using a gradient (5 to 30%) ofacetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of 34-2 (118 mg). [M+H]⁺ calcd for C₂₅H₂₄N₈O₂ 468.52, found 469.0.

Step B: Preparation of azetidin-3-ylmethyl1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzyl)azetidine-3-carboxylate(88). A solution of 34-2 (23 mg, 0.033 mmol), 33-5 (9.27 mg, 0.050mmol), HATU (18.83 mg, 0.050 mmol), DMAP (0.40 mg, 3.3 μmol), and DIEA(23.0 μl, 0.132 mmol) in DCM (0.5 ml) was stirred at 25° C. for 16 h.TFA (127 μl) was added to the reaction mixture and was heated at 45° C.for 30 min. The reaction mixture was concentrated in vacuum and waspurified by preparative HPLC chromatography using a gradient (2 to 40%)of acetonitrile in water with 0.05% trifluoroacetic acid to yield a TFAsalt of the title compound (24.0 mg). [M+H]⁺ calcd for C₂₉H₃₁N₉O₂537.63, found 538.2.

Example 35: Synthesis of (S)-1-methylpyrrolidin-3-yl(R)-1-((2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazol-4-yl)methyl)piperidine-3-carboxylate(508)

Step A: Preparation of methyl2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-4-carboxylate(35-2). To a suspension of 1-8 (3.6 g, 12.2 mmol) in 1,4-dioxane (100ml) was added 35-1 (2.5 g, 15.8 mmol), Cs₂CO₃ (8.0 g, 24.4 mmol),Pd₂(dba)₃ (1.1 g, 1.22 mmol) and BINAP (760 mg, 1.22 mmol). The reactionmixture was degassed under vacuum and N₂ was purged for 3 times. Thereaction mixture was heated at 110° C. for 12 h. The reaction mixturewas concentrated in vacuum to give the residue. The residue was purifiedby preparative HPLC chromatography using a gradient (12 to 42%) ofacetonitrile in water with 0.1% trifluoroacetic acid to yield a TFA saltof 35-2 (1.7 g). [M+H]⁺ calcd for C₁₉H₁₆N₈O₂S 420.45, found 421.0.

Step B: Preparation of(2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazol-4-yl)methanol(35-3). To a solution of 35-2 (50 mg×16, 0.119 mmol×16) in THF (1 mL×16)was added LAH (9 mg×16, 0.238 mmol×16) at 0° C. The mixture was stirredat 25° C. for 20 min. The reaction mixture was diluted with Rochellesalt solution (100 ml) and extracted with DCM (200 ml×3), The organiclayer was dried over Na₂SO₄, filtered, and concentrated in vacuum. Theresidue was purified by preparative HPLC chromatography using a gradient(10 to 35%) of acetonitrile in water with 0.225% formic acid to obtain35-3 (86 mg) as yellow solid. [M+H]⁺ calcd for C₁₈H₁₆N₈OS 392.44, found393.1.

Step C: Preparation ofN2-(4-(chloromethyl)thiazol-2-yl)-N4-(2-(6-methylpyridin-2-yl)pyrimidin-4-yl)pyrimidine-2,4-diamine(35-4). To a solution of 35-3 (101 mg, 0.257 mmol) in DCM (6 ml) wasadded SOCl₂ (92 mg, 0.771 mmol) dropwise at 0° C. The reaction mixturewas stirred at 25° C. for 2 h. The reaction mixture was concentrated invacuum. The residue was purified by trituration to give 35-4 (50 mg) asyellow solid. [M+H]⁺ calcd for C₁₈H₁₅ClN₈OS 410.88, found 411.0.

Step D: Preparation of(R)-1-((2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazol-4-yl)methyl)piperidine-3-carboxylicacid (35-6). To a solution of 35-4 (200 mg, 0.487 mmol), 35-5 (162 mg,0.730 mmol), and DIEA (425 μl, 2.434 mmol) in DCM (7.8 ml) was heated at55° C. for 2 h. TFA (938 μl) was added to the reaction mixture and washeated at 45° C. for 30 min. The reaction mixture was concentrated invacuum. The residue was purified by preparative HPLC chromatographyusing a gradient (5 to 75%) of acetonitrile in water with 0.05%trifluoroacetic acid to yield a TFA salt of 35-6 (253 mg). [M+H]⁺ calcdfor C₂₄H₂₅N₉O₂S 503.59, found 504.0.

Step E: Preparation of (S)-1-methylpyrrolidin-3-yl(R)-1-((2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazol-4-yl)methyl)piperidine-3-carboxylate(508). A solution of 35-6 (25 mg, 0.034 mmol), 35-7 (5.18 mg, 0.051mmol), EDCI (9.83 mg, 0.051 mmol), and DMAP (0.84 mg, 6.83 μmol) in DCM(0.5 ml) was stirred at 25° C. for 16 h. TFA (132 μl) was added to thereaction mixture and was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (2 to 50%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(18.2 mg). [M+H]⁺ calcd for C₂₉H₃₄N₁₀O₂S 586.72, found 587.1.

Example 36: Synthesis of (R)-1-methylpyrrolidiVn-3-yl2-(1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazin-2-yl)acetate(103)

Step A: Preparation of2-(4-(tert-butoxycarbonyl)-1-(4-nitrophenyl)piperazin-2-yl)acetic acid(36-2). To a solution of 5-1 (1.0 g, 7.09 mmol) and 36-1 (2.195 g, 8.51mmol) in ACN (12 ml) and water (3 ml) was added potassium carbonate (4.9g, 35.46 mmol), followed by 18-crown-6 (93 mg, 0.354 mmol). The reactionmixture was heated at 90° C. for 48 h. The reaction mixture wasconcentrated and the crude mixture diluted with water (10 ml). Theaqueous layer was washed with EtOAc and the pH of aqueous layer wasadjusted to ˜4 using saturated citric acid solution and extracted withDCM (5 ml×2). The organic layer was concentrated to get the cruderesidue of 36-2 (750 mg) as a yellow solid. [M+H]⁺ calcd for C₁₇H₂₃N₃O₆365.39, found 366.22.

Step B: Preparation of tert-butyl3-(2-methoxy-2-oxoethyl)-4-(4-nitrophenyl)piperazine-1-carboxylate(36-3). To a solution of 36-2 (700 mg, 1.91 mmol) in ACN (7 ml) and MeOH(3 ml) was added TMS-CHN₂ in hexane (2M) (5.8 mL 11.5 mmol) and thereaction mixture was stirred at 25° C. for 3 h. To the reaction mixturewas added excess methanol and stirred at 25° C. for 15 min, thenresidual solvent was distilled off to obtain the crude residue. Theresidue was purified by silica-gel column chromatography using agradient (0 to 15%) of EtOAc in Hexane to give 36-3 (700 mg) as yellowsolid. [M+H]⁺ calcd for C₁₈H₂₅N₃O₆ 379.41, found 380.03.

Step C: Preparation of tert-butyl4-(4-aminophenyl)-3-(2-methoxy-2-oxoethyl)piperazine-1-carboxylate(36-4). To a solution of 36-3 (1.3 g, 3.43 mmol) in THF:IPA (20 ml, 1:1)was added 10% Pd/C (1.3 g), then the resulting suspension was stirred at25° C. under H₂ atmosphere for 3 h. The reaction mixture was filteredthrough Celite bed and the residue was washed with EtOAc. The organiclayer was then evaporated under reduced pressure to get the crudeproduct 36-4 (1.3 g), which was used directly for the next step. [M+H]⁺calcd for C₁₈H₂₇N₃O₄ 349.43, found 350.10.

Step D: Preparation of tert-butyl3-(2-methoxy-2-oxoethyl)-4-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazine-1-carboxylate(36-5). To a stirred solution of 1-8 (1.2 g, 4.02 mmol) and 36-4 (1.6 g,4.83 mmol) in 1,4-dioxane (15 ml) was added Cs₂CO₃ (2.62 g, 8.05 mmol)and the reaction mixture was purged with N₂ for 30 min. To the reactionmixture was added Brettphos Pd G4 (0.370 g, 0.40 mmol), then purging wasrepeated with N2 for 5 min. The reaction mixture was heated at 110° C.for 3 h. The reaction mixture was filtered through a Celite bed, washedwith 5% MeOH:DCM, and concentrated to afford a crude residue. Theresidue was purified by silica-gel column chromatography using agradient (0 to 100%) of EtOAc in PE to give 36-5 (1.18 g) as a whitesolid.

Step E: Preparation of2-(4-(tert-butoxycarbonyl)-1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazin-2-yl)aceticacid (36-6). To a stirred solution of 36-5 (1.4 g, 2.29 mmol) in MeOH(10 ml), THF (5 ml) and water (2.5 ml) was added LiOH·H₂O (0.48 g, 11.45mmol) and the reaction mixture was stirred at 25° C. for 16 h. Thereaction mixture pH was adjusted to ˜3-4 by using aq. citric acidsolution. Desired acid was extracted from the aqueous layer using 10%MeOH:DCM. The organic layer was dried over Na₂SO₄, filtered andconcentrated to get crude residue. The residue was purified bysilica-gel column chromatography using a gradient (5 to 10%) of MeOH inDCM to give 36-6 (1.12 g) as an off-white solid. [M+H]⁺ calcd forC₃₁H₃₅N₉O₄ 597.68, found 598.32.

Step F: Preparation of (R)-1-methylpyrrolidin-3-yl2-(1-(4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)phenyl)piperazin-2-yl)acetate(103). A solution of 36-6 (30 mg, 0.050 mmol), 36-7 (7.62 mg, 0.075mmol), EDCI (14.43 mg, 0.075 mmol), and DMAP (1.20 mg, 10.0 μmol) in DCM(0.5 ml) was stirred at 25° C. for 16 h. TFA (193 μl) was added to thereaction mixture and was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (2 to 50%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(11.4 mg). [M+H]⁺ calcd for C₃₁H₃₆N₁₀O₂ 580.70, found 581.1.

Example 37: Synthesis of azetidin-3-ylmethyl4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-2-carboxylate(96)

A solution of 30-1 (50 mg, 0.123 mmol), 33-5 (34.6 mg, 0.185 mmol), EDCI(35.5 mg, 0.185 mmol), and DMAP (3.01 mg, 0.025 mmol) in DCM (0.5 ml)was stirred at 25° C. for 16 h. TFA (475 μl) was added to the reactionmixture and was heated at 45° C. for 30 min. The reaction mixture wasconcentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (2 to 30%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(22.8 mg). [M+H]⁺ calcd for C₂₃H₂₂N₈O₂S 474.54, found 475.1.

Example 38: Synthesis of azetidin-3-ylmethyl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-3-carboxylate(142)

Step A: Preparation of methyl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-3-carboxylate(38-2). A solution of 1-8 (1.6 g, 5.36 mmol), 38-1 (842 mg, 5.36 mmol)and conc. HCl (0.6 ml) in IPA (16 ml) was heated at 80° C. for 12 h. Thereaction mixture was then allowed to cool to 25° C., diluted with IPAand filtered to obtain the HCl salt of 38-2 (900 mg) as an off-whitesolid. [M+H]⁺ calcd for C₂₀H₁₇N₇O₂S 419.46, found 420.17.

Step B: Preparation of5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-3-carboxylicacid (38-3). To a stirred solution of 38-2 (900 mg, 2.14 mmol) in MeOH(6 ml), THF (4 ml) and water (2 ml) was added KOH (240 mg, 4.29 mmol)and the resulting mixture stirred and heated at 65° C. for 3 h. Thereaction mixture was concentrated and diluted with water and acidifiedwith 3 N HCl to (pH 4-5) until light brown solid fell out, which wasfiltered and washed with acetonitrile and diethyl ether to get thedesired compound 38-3 (215 mg) as a light brown solid. [M+H]⁺ calcd forC₁₉H₁₅N₇O₂S 405.44, found 406.03.

Step C: Preparation of azetidin-3-ylmethyl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiophene-3-carboxylate(142). A solution of 38-3 (25 mg, 0.062 mmol), 33-5 (17.32 mg, 0.092mmol), EDCI (17.73 mg, 0.092 mmol), and DMAP (1.50 mg, 0.012 mmol) inDCM (0.5 ml) was stirred at 25° C. for 16 h. TFA (238 μl) was added tothe reaction mixture and was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (5 to 50%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(17.3 mg). [M+H]⁺ calcd for C₂₃H₂₂N₈O₂S 474.54, found 475.1.

Example 39: Synthesis of piperidin-4-yl2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-4-carboxylate(148)

Step A: Preparation of2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-4-carboxylicacid (39-1). A suspension of 35-2 (1.0 g, 2.38 mmol) and NaOH (1.9 g,47.6 mmol) in MeOH (30 ml) and H₂O (30 ml) was heated at 60° C. for 5 h.The reaction mixture was concentrated in vacuum to remove MeOH andacidified by 2M HCl to pH=6-7. The reaction mixture was filtered. Thecake was washed with H₂O (50 ml) and lyophilized to obtain 39-1 (600 mg)as yellow solid. [M+H]⁺ calcd for C₁₈H₁₄N₈O₂S 406.42, found 407.0.

Step B: Preparation of piperidin-4-yl2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-4-carboxylate(148). A solution of 39-1 (25 mg, 0.062 mmol), 39-2 (18.57 mg, 0.092mmol), EDCI (17.69 mg, 0.092 mmol), and DMAP (1.50 mg, 0.012 mmol) inDCM (0.5 ml) was stirred at 25° C. for 16 h. TFA (351 μl) was added andthe reaction mixture was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (5 to 65%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(9.2 mg). [M+H]⁺ calcd for C₂₃H₂₃N₉O₂S 489.56, found 490.0.

Example 40: Synthesis of azetidin-3-ylmethyl2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-5-carboxylate(338)

Step A: Preparation of ethyl2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-5-carboxylate(40-2). To a stirred solution of 1-8 (600 mg, 2.013 mmol) and 40-1 (519mg, 3.019 mmol) in 1,4-dioxane (12 ml) was added cesium carbonate (1.3g, 4.026 mmol) and purged for 15 min. Pd2(dba)3 (184 mg, 0.201 mmol) andBINAP (250 mg, 0.402 mmol) were then added. The reaction mixture wasthen heated at 110° C. for 3 h. The reaction mixture was diluted with10% MeOH in DCM, filtered through Celite bed and concentrated to getcrude residue. The residue was purified by silica-gel columnchromatography using a gradient (0 to 5%) of MeOH in DCM to give 40-2(430 mg). [M+H]⁺ calcd for C₂₀H₁₈N₈O₂S 434.48, found 435.10.

Step B: Preparation of2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-5-carboxylicacid (40-3). To the solution of 40-2 (300 mg, 0.69 mmol) in1,4-dioxane:water (9.0:0.5 ml) was added 6N HCl (1.5 ml). The reactionmixture was heated at 90° C. for 12 h. The reaction mixture wasconcentrated completely to dryness. The resulting residue was trituratedusing MeOH, DCM, diethyl ether and ACN to get desired compound which waspurified further through preparative HPLC chromatography to obtain 40-3(70 mg). [M+H]⁺ calcd for C₁₈H₁₄N₈O₂S 406.42, found 407.18.

Step C: Preparation of azetidin-3-ylmethyl2-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)thiazole-5-carboxylate(338). A solution of 40-3 (15 mg, 0.037 mmol), 33-5 (10.37 mg, 0.055mmol), EDCI (10.61 mg, 0.055 mmol), and DMAP (0.90 mg, 7.38 μmol) in DCM(0.5 ml) was stirred at 25° C. for 16 h. TFA (142 μl) was added to thereaction mixture and was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (5 to 50%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(5.7 mg). [M+H]⁺ calcd for C₂₅H₂₄N₈O₂S 475.53, found 476.0.

Example 41: Synthesis of azetidin-3-ylmethyl3-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoate(178)

Step A: Preparation of methyl3-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoate(41-2). To a mixture of 1-8 (600 mg, 2.01 mmol) and 41-1 (304 mg, 2.01mmol) in IPA (10 ml) was added HCl (0.5 mL). The reaction mixture washeated at 80° C. for 16 h. The reaction mixture was quenched with H₂O(30 ml) and filtered. The filter cake was evaporated to dryness to give41-2 (820 mg) as white solid. [M+H]⁺ calcd for C₂₂H₁₉N₇O₂ 413.44, found414.2.

Step B: Preparation of3-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoicacid (41-3). A mixture of 41-2 (780 mg, 1.87 mmol) in EtOH (10 ml) andaq. NaOH (10 ml) was heated at 80° C. for 16 h. The reaction mixture wasconcentrated in vacuum to remove EtOH. To the residue was added HCl toadjust the pH to 5, then the mixture was filtered. The filter cake waswashed with H₂O (30 ml) and the residue was evaporated to dryness togive 41-3 (530 mg) as yellow solid. [M+H]⁺ calcd for C₂₁H₁₇N₇O₂ 399.41,found 399.9.

Step C: Preparation of azetidin-3-ylmethyl3-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoate(178). A solution of 41-3 (25 mg, 0.063 mmol), 33-5 (17.58 mg, 0.094mmol), EDCI (18.00 mg, 0.094 mmol), and DMAP (1.53 mg, 0.013 mmol) inDCM (0.5 ml) was stirred at 25° C. for 16 h. TFA (240 μl) was added tothe reaction mixture and was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (5 to 50%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(29.5 mg). [M+H]⁺ calcd for C₂₅H₂₄N₈O₂ 468.52, found 469.1.

Example 42: Synthesis of azetidin-3-ylmethyl4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoate(33)

Step A: Preparation of4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoicacid (42-1). A mixture of 32-2 (780 mg, 1.87 mmol) in EtOH (10 ml) andaq. NaOH (10 ml) was stirred at 80° C. for 16 h. The reaction mixturewas concentrated in vacuum to remove EtOH. To the residue was added HClto adjust the pH to 5, then the mixture was filtered. The filter cakewas washed with H₂O (30 ml) and the residue evaporated to dryness togive 42-1 (530 mg) as yellow solid. [M+H]⁺ calcd for C₂₁H₁₇N₇O₂ 399.41,found 399.9.

Step B: Preparation of azetidin-3-ylmethyl4-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)benzoate(33). A solution of 42-1 (25 mg, 0.063 mmol), 33-5 (17.58 mg, 0.094mmol), EDCI (18.00 mg, 0.094 mmol), and DMAP (1.53 mg, 0.013 mmol) inDCM (0.5 ml) was stirred at 25° C. for 16 h. TFA (240 μl) was added tothe reaction mixture and was heated at 45° C. for 30 min. The reactionmixture was concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (5 to 65%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(21.1 mg). [M+H]⁺ calcd for C₂₅H₂₄N₈O₂ 468.52, found 469.1.

Example 43: Synthesis of azetidin-3-ylmethyl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)picolinate(545)

Step A: Preparation of methyl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)picolinate(43-2). A vial of 1-8 (250 mg, 0.837 mmol), 44-1 (127 mg, 0.837 mmol),cesium carbonate (545 mg, 1.674 mmol), BrettPhos (44.9 mg, 0.084 mmol),and BrettPhos Pd G4 (77 mg, 0.084 mmol) in degassed 1,4-dioxane (4.2 ml)was heated to 110° C. for 16 h. The reaction mixture was concentrated invacuum and purified by silica-gel column chromatography using a gradient(0 to 15%) of MeOH in DCM to obtain 43-2 (164 mg) as an orange oil.[M+H]⁺ calcd for C₂₁H₁₈N₈O₂ 414.43, found 415.

Step B: Preparation of5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)picolinicacid (43-3). A solution of 43-2 (164 mg, 0.396 mmol), and LiOH (24.73mg, 1.033 mmol) in a 3:2:1 mixture of THF (1291 μl), MeOH (861 μl), andH₂O (430 μl) was heated to 65° C. for 3 h. The reaction mixture wasconcentrated in vacuum and purified by preparative HPLC chromatographyusing a gradient (5 to 30%) of acetonitrile in water with 0.05%trifluoroacetic acid to yield a TFA salt of 43-3 (129 mg). [M+H]⁺ calcdfor C₂₀H₁₆N₈O₂ 400.40, found 401.1.

Step C: Preparation of azetidin-3-ylmethyl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)picolinate(545). A solution of 43-3 (20 mg, 0.039 mmol), 33-5 (10.92 mg, 0.058mmol), HATU (22.17 mg, 0.058 mmol), and DIEA (27.2 μl, 0.156 mmol) inDCM (0.5 ml) was heated to 50° C. for 1 h. TFA (120 μl) was added to thereaction mixture and stirred at 25° C. for 30 min. The reaction mixturewas concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (2 to 60%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(22.8 mg). [M+H]⁺ calcd for C₂₄H₂₃N₉O₂ 469.51, found 470.1.

Example 44: Synthesis of (R)-pyrrolidin-3-yl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)nicotinate(549)

Step A: Preparation of methyl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)nicotinate(44-2). A vial of 1-8 (250 mg, 0.837 mmol), 44-1 (127 mg, 0.837 mmol),cesium carbonate (545 mg, 1.674 mmol), BrettPhos (44.9 mg, 0.084 mmol),and BrettPhos Pd G4 (77 mg, 0.084 mmol) in degassed 1,4-dioxane (4.2 ml)was heated to 110° C. for 16 h. The reaction mixture was concentrated invacuum and purified by silica-gel column chromatography using a gradient(0 to 15%) of MeOH in DCM to obtain 44-2 (214 mg) as an orange oil.[M+H]⁺ calcd for C₂₁H₁₈N₈O₂ 414.43, found 415.

Step B: Preparation of5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)nicotinicacid (44-3). A solution of 44-2 (214 mg, 0.516 mmol) and LiOH (24.73 mg,1.033 mmol) in a 3:2:1 mixture of THF (1291 μl), MeOH (861 μl), and H₂O(430 μl) was heated to 65° C. for 3 h. The reaction mixture wasconcentrated in vacuum and purified by preparative HPLC chromatographyusing a gradient (5 to 30%) of acetonitrile in water with 0.05%trifluoroacetic acid to yield a TFA salt of 44-3 (77 mg). [M+H]⁺ calcdfor C₂₀H₁₆N₈O₂ 400.40, found 401.1.

Step C: Preparation of (R)-pyrrolidin-3-yl5-((4-((2-(6-methylpyridin-2-yl)pyrimidin-4-yl)amino)pyrimidin-2-yl)amino)nicotinate(549). A solution of 44-3 (20 mg, 0.039 mmol), 44-4 (10.92 mg, 0.058mmol), HATU (22.17 mg, 0.058 mmol), and DIEA (27.2 μl, 0.156 mmol) inDCM (0.5 ml) was heated to 50° C. for 1 h. TFA (120 μl) was added to thereaction mixture and stirred at 25° C. for 30 min. The reaction mixturewas concentrated in vacuum and was purified by preparative HPLCchromatography using a gradient (2 to 60%) of acetonitrile in water with0.05% trifluoroacetic acid to yield a TFA salt of the title compound(12.8 mg). [M+H]⁺ calcd for C₂₄H₂₃N₉O₂ 469.51, found 470.1.

Example 45: Biochemical ALK5 (TGF-βR1) Assay to Measure pKi

Apparent pKi values for compounds of the present disclosure weredetermined using a recombinant human ALK5 (TGF-βR1) protein (Product No.PR9075A or equivalent, Life Technologies) and a commercially-availablekinase assay (LANCE® (lanthanide chelate excite) Ultra ULight™ kinaseassay, Product Nos. TRF0130-M and TRF02108-M, Perkin Elmer) as describedbelow.

The assays were performed in a 384-well plate (24 columns×16wells/rows). An Echo®550 Liquid Handler (Labcyte) was used to preparevarious intermediate concentrations of compounds of the presentdisclosure in 100% DMSO. From the intermediate concentrations, a rangeof concentrations (from 10 μM to 25 μM corresponding to volumes up to105 nL) were prepared and ejected into a final assay plate to be used tocreate individual dose response curves for each of the subjectcompounds. To a separate column within the assay plate, 105 nL of DMSOin each well was used to establish a maximum assay signal. Additionally,105 nL of 100 μM SD-208, a selective TGF-βR1 inhibitor (Catalog #S7624,Selleck Chemicals), was used in another column of wells to establish aminimal assay signal.

With a multidrop dispenser, 8 μL of enzyme mixture (1.25× final) wasadded to each well. The enzyme mixture consisted of 250 μM ALK5 enzymeand 62.5 nM peptide substrate (LANCE® (lanthanide chelate excite) UltraULight™-DNA Topoisomerase 2-alpha (Thr1342)) prepared in assay buffer(50 mM HEPES, 10 mM MgCl₂, 1 mM EGTA, 0.01% Tween-20, pH 7.5 at roomtemperature) with 2 mM DTT added prior to use. The plate was then sealedwith an adhesive seal and allowed to equilibrate for 60 minutes at roomtemperature.

Next, 2 μL of 125 μM ATP (5×final, 125 μM ATP prepared in assay bufferwith 2 mM DTT) was added to the incubated mixtures, covered with aMicroClime® Environmental Lid (Product No. LLS-0310, Labcyte) andimmediately transferred to 37° C. The reactions were allowed to proceedat 37° C. for 60 minutes before terminating with the addition of 10 μLof detection antibody (LANCE® (lanthanide chelate excite) UltraEuropium-anti-phospo-DNA Topoisomerase 2-alpha (Thr1342)) in detectionmixture (12 mM EDTA, 4 nM detection antibody prepared in detectionbuffer (50 mM Tris-HCl, 150 mM NaCl, 0.5% BSA (Fraction V), pH 7.0)) atroom temperature. The plate was then read on a Perkin Elmer EnVisionPlate Reader using europium specific reader settings with excitation andemission wavelengths set to 320 or 340 nm and 665 nm, respectively.These data were used to calculate percent enzyme inhibition values basedon DMSO and SD-208 background controls.

For dose-response analyses, percent inhibition versus compoundconcentrations were plotted, and pIC₅₀ values were determined from a4-parameter robust fit model with GraphPad Prism V5 Software (GraphPadSoftware, Inc., La Jolla, CA). This model obtains pIC₅₀ values byfitting the sigmoidal dose-response (variable slope) equation to thedata. Results were expressed as pIC₅₀ (negative logarithm of IC₅₀) andsubsequently converted to pK_(i) (negative logarithm of dissociateconstant, K_(i)) using the Cheng-Prusoff equation. The higher the valueof pK_(i) (lower value of K_(i)), the greater the inhibition of ALK5activity. Certain compounds disclosed herein exhibited pK_(i) values ofgreater than 8 or greater than 9 when tested in the biochemical ALK5assay.

Table 3 shows biological activities of selected compounds in abiochemical ALK5 assay. Compound numbers correspond to the numbers andstructures provided in Tables 1 and 2 and Examples 1-44.

TABLE 3 7.5 to 8.4 (+) 8.5 to 9.4 (++) 9.5 to 10.4 (+++) ≥10.5 (++++)ALK5 19, 30, 35, 87, 2, 3, 5, 16, 18, 1, 4, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 17, 24, 29, 42, 46, pK_(i) 119, 165, 166, 20, 26, 36, 43, 21,22, 23, 25, 27, 28, 31, 32, 33, 34, 37, 89, 94, 99, 226, 234, 306, 47,54, 56, 62, 38, 39, 40, 41, 44, 45, 48, 49, 50, 51, 52, 123, 144, 153,309, 325, 333, 66, 72, 82, 86, 53, 55, 57, 58, 59, 60, 61, 63, 64, 65,67, 175, 188, 189, 338, 348, 399, 90, 91, 103, 68, 69, 70, 71, 73, 74,75, 76, 77, 78, 79, 208, 223, 252, 410, 426, 468, 107, 114, 117, 80, 81,83, 84, 85, 88, 92, 93, 95, 96, 97, 261, 319, 327, 502, 526, 529, 121,126, 146, 98, 100, 101, 102, 104, 105, 106, 108, 331, 336, 343, 535,564, 574, 148, 155, 162, 109, 110, 111, 112, 113, 115, 116, 118, 364,373, 378, 575, 578, 579, 169, 170, 171, 120, 122, 124, 125, 127, 128,129, 130, 385, 404, 405, 581, 585, 592, 176, 178, 179, 131, 132, 133,134, 135, 136, 137, 138, 474, 503, 539 601, 604, 608, 191, 201, 230,139, 140, 141, 142, 143, 145, 147, 149, 622, 624, 630, 236, 250, 255,150, 151, 152, 154, 156, 157, 158, 159, 632, 633, 634, 256, 257, 267,160, 161, 163, 164, 167, 168, 172, 173, 636, 640, 281, 284, 285, 174,177, 180, 181, 182, 183, 184, 185, 1000, 1003, 287, 291, 292, 186, 187,190, 192, 193, 194, 195, 196, 1004, 1008, 296, 303, 304, 197, 198, 199,200, 202, 203, 204, 205, 1020, 1043 308, 310, 316, 206, 207, 209, 210,211, 212, 213, 214, 321, 323, 329, 215, 216, 217, 218, 219, 220, 221,222, 339, 349, 350, 224, 225, 227, 228, 229, 231, 232, 233, 354, 355,363, 235, 237, 238, 239, 240, 241, 242, 243, 369, 371, 379, 244, 245,246, 247, 248, 249, 251, 253, 386, 388, 391, 254, 258, 259, 260, 262,263, 264, 265, 396, 401, 402, 266, 268, 269, 270, 271, 272, 273, 274,406, 408, 413, 275, 276, 277, 278, 279, 280, 282, 283, 423, 425, 437,286, 288, 289, 290, 293, 294, 295, 297, 439, 442, 446, 298, 299, 300,301, 302, 305, 307, 311, 455, 459, 461, 312, 313, 314, 315, 317, 318,320, 322, 467, 469, 471, 324, 326, 328, 330, 332, 334, 335, 337, 481,485, 492, 340, 341, 342, 344, 345, 346, 347, 351, 498, 500, 505, 352,353, 356, 357, 358, 359, 360, 361, 508, 511, 512, 362, 365, 366, 367,368, 370, 372, 374, 513, 514, 517, 375, 376, 377, 380, 381, 382, 383,384, 525, 527, 532, 387, 389, 390, 392, 393, 394, 395, 397, 548, 549,550, 398, 400, 403, 407, 409, 411, 412, 414, 551, 553, 570, 415, 416,417, 418, 419, 420, 421, 422, 580, 586, 589, 424, 427, 428, 429, 430,431, 432, 433, 593, 595, 606, 434, 435, 436, 438, 440, 441, 443, 444,609, 611, 613, 445, 447, 448, 449, 450, 451, 452, 453, 615, 616, 618,454, 456, 457, 458, 460, 462, 463, 464, 623, 641, 465, 466, 470, 472,473, 475, 476, 477, 1001, 1006, 478, 479, 480, 482, 483, 484, 486, 487,1007, 1009, 488, 489, 490, 491, 493, 494, 495, 496, 1017, 1019, 497,499, 501, 504, 506, 507, 509, 510, 1021, 1022, 515, 516, 518, 519, 520,521, 522, 523, 1023, 1044, 524, 528, 530, 531, 533, 534, 536, 537, 1045538, 540, 541, 542, 543, 544, 545, 546, 547, 552, 554, 556, 557, 558,560, 561, 562, 563, 565, 566, 567, 568, 569, 571, 572, 573, 576, 577,582, 583, 584, 587, 588, 590, 591, 594, 596, 597, 598, 599, 600, 602,603, 605, 607, 610, 612, 614, 617, 619, 620, 621, 625, 626, 627, 628,629, 631, 635, 637, 638, 639, 642, 1002, 1005, 1010, 1011, 1012, 1013,1014, 1015, 1016, 1018, 1024, 1025, 1026, 1027, 1028, 1029, 1030, 1031,1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042

Example 46: Cellular ALK5 Potency Assay to Measure pIC₅₀, Inhibition ofTGF-β Stimulated pSMAD3 Formation in BEAS-2B Cells

The potency of compounds of the present disclosure for inhibition ofTGF-β-stimulated SMAD3 phosphorylation was measured in BEAS-2B cells, ahuman lung epithelial cell line. TGF-β signals through activinreceptor-like kinase 5 (ALK5) immediately prior to SMAD3phosphorylation. As the AlphaLISA SureFire Ultra kit (Perkin Elmer)quantitatively measures pSMAD3 levels in lysate, the assay demonstratesthe ALK5 cellular potency of a test compound.

BEAS-2B cells were grown using 50% DMEM (Life Technologies) and 50% F-12(Life Technologies) media, supplemented with 10% Fetal Bovine Serum(ATCC), 25 mM HEPES (Life Technologies), and 1×Pen-Strep (LifeTechnologies). Cells were cultured in a humidified incubator set at 37°C., 5% CO₂, and trypsonized using 0.25% Trypsin with 0.5%polyvinylpyrrolidone (PVP).

For the assay, BEAS-2B cells were seeded at 7,500 cells/well (25μL/well) in a 384-well plate and cultured overnight. Before dosing,growth media was aspirated and the wells were rinsed with HBSS Buffer(HBSS with Calcium and Magnesium, Life Technologies) supplemented with25 mM HEPES (Life Technologies) and 1% Bovine Serum Albumin (Roche).Compounds were serially diluted in DMSO, then further diluted withsupplemented HBSS Buffer (50 μL/well) to create a compound plate 3× ofthe final assay concentration, at 0.3% DMSO. The diluted compounds werethen added to the cells (8 μL/well) and incubated at 37° C., 5% CO₂ for1 hour. After the compound incubation, TGF-β (R&D Systems) reconstitutedin supplemented HBSS Buffer was added to the cells (12 μL/well, finalconcentration 10 ng/mL) and incubated for a further 30 minutes, afterwhich the cells were immediately lysed with AlphaLISA lysis buffer(PerkinElmer). AlphaLISA Acceptor and Detector beads (PerkinElmer) wereadded 2 hours apart, then incubated overnight to be read the next day.The potency of the compound was determined through analysis ofdose-dependent quantified changes in pSMAD3 signal from baseline(non-compound treated TGF-β stimulated cells). Data are expressed aspIC₅₀ (negative decadic logarithm IC₅₀) values. Certain compoundsdisclosed herein exhibited pIC₅₀ values of greater than 6 or greaterthan 7 when tested in BEAS-2B3 cells.

Table 4 shows biological activities of selected compounds in a cellularALK5 potency assay. Compound numbers correspond to the numbers andstructures provided in Tables 1 and 2 and Examples 1-44.

TABLE 4 5 to 5.8 (+) 5.9 to 6.7 (++) 6.8 to 7.6 (+++) ≥7.7 (++++) BEAS2B26, 82, 386, 11, 30, 31, 37, 1, 2, 3, 4, 6, 7, 8, 9, 12, 13, 15, 16, 17,5, 10, 14, 20, pIC₅₀ 423, 540, 603, 48, 53, 57, 68, 18, 19, 21, 22, 23,24, 25, 27, 32, 33, 34, 28, 29, 43, 45, 610, 629, 636, 86, 87, 91, 93,35, 36, 38, 39, 40, 41, 42, 44, 46, 47, 49, 58, 76, 78, 89, 1012, 1014,95, 103, 107, 50, 51, 52, 54, 55, 56, 59, 60, 61, 62, 63, 92, 94, 126,1044 114, 117, 155, 64, 65, 66, 67, 69, 70, 71, 72, 73, 74, 75, 128,135, 139, 157, 159, 162, 77, 79, 80, 81, 83, 84, 85, 88, 90, 96, 97,161, 193, 203, 181, 200, 206, 98, 99, 100, 101, 102, 104, 105, 106, 208,209, 210, 215, 224, 237, 108, 109, 110, 111, 112, 113, 115, 116, 227,235, 254, 245, 251, 256, 118, 119, 120, 121, 122, 123, 124, 125, 261,265, 268, 269, 283, 286, 127, 129, 130, 131, 132, 133, 134, 136, 284,288, 294, 296, 297, 308, 137, 138, 140, 141, 142, 143, 144, 145, 315,328, 331, 309, 326, 338, 146, 147, 148, 149, 150, 151, 152, 153, 351,364, 365, 339, 340, 348, 154, 156, 158, 160, 163, 164, 165, 166, 387,388, 421, 349, 358, 363, 167, 168, 169, 170, 171, 172, 173, 174, 443,452, 465, 373, 379, 390, 175, 176, 177, 178, 179, 180, 182, 183, 475,477, 514, 394, 402, 406, 184, 185, 186, 187, 188, 189, 190, 191, 523,535, 571 413, 416, 422, 192, 194, 195, 196, 197, 198, 199, 201, 428,429, 439, 202, 204, 205, 207, 211, 212, 213, 214, 450, 454, 455, 216,217, 218, 219, 220, 221, 222, 223, 468, 471, 473, 225, 226, 228, 229,230, 231, 232, 233, 478, 483, 484, 234, 236, 238, 239, 240, 241, 242,243, 488, 497, 498, 244, 246, 247, 248, 249, 250, 252, 253, 499, 517,530, 255, 257, 258, 259, 260, 262, 263, 264, 532, 551, 557, 267, 270,271, 272, 273, 274, 275, 276, 562, 563, 564, 277, 278, 279, 280, 281,282, 285, 287, 569, 574, 575, 289, 290, 291, 292, 293, 295, 298, 299,578, 580, 585, 300, 301, 302, 303, 304, 305, 306, 307, 594, 596, 601,310, 311, 312, 313, 314, 316, 317, 318, 604, 605, 608, 320, 321, 322,323, 324, 325, 327, 329, 611, 615, 624, 330, 332, 333, 334, 335, 336,337, 341, 630, 633, 342, 343, 344, 345, 346, 347, 350, 352, 1001, 1002,353, 354, 355, 356, 357, 359, 360, 361, 1003, 1006, 362, 366, 367, 368,369, 370, 371, 372, 1007, 1008, 374, 375, 376, 377, 378, 380, 381, 382,1009, 1010, 383, 384, 385, 389, 391, 392, 393, 395, 1013, 1017, 396,397, 398, 399, 400, 401, 403, 404, 1019, 1020, 405, 407, 408, 409, 410,411, 412, 414, 1022, 1023, 415, 417, 418, 419, 420, 424, 425, 426, 1043427, 430, 431, 432, 433, 434, 435, 436, 437, 438, 440, 441, 442, 444,445, 446, 447, 448, 449, 451, 453, 456, 457, 458, 459, 460, 461, 462,463, 464, 466, 467, 469, 470, 472, 474, 476, 479, 480, 481, 482, 485,486, 487, 489, 490, 491, 492, 493, 494, 495, 496, 500, 501, 502, 503,504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 515, 516, 518, 519,520, 521, 522, 524, 525, 526, 527, 528, 529, 531, 533, 534, 536, 537,538, 539, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 552, 553,554, 556, 558, 560, 561, 565, 566, 567, 568, 570, 572, 573, 576, 577,579, 581, 582, 583, 584, 586, 587, 588, 589, 590, 591, 592, 593, 595,597, 598, 599, 600, 602, 606, 607, 609, 612, 613, 614, 616, 617, 618,619, 620, 621, 622, 623, 625, 626, 627, 628, 631, 632, 634, 635, 637,638, 639, 641, 642, 1000, 1004, 1005, 1011, 1015, 1016, 1018, 1021,1024, 1025, 1026, 1027, 1028, 1029, 1030, 1031, 1032, 1033, 1034, 1035,1036, 1037, 1038, 1039, 1040, 1041, 1042, 1045

Example 47: Cytotoxicity Measured by Premature Chromosome Condensation[15] (pCC₁₅)

The impact of a compound of the present disclosure on cellular adenosinetriphosphate (ATP) levels was measured in Beas2B cells, a human lungepithelial cell line. Levels of ATP are correlated with the viability ofcells and are often measured to determine the potential cytotoxicity ofcompounds. CellTiter-Glo, which lyses the cells and produces aluminescent signal proportional to the amount of ATP present, was usedto determine the effect of test compound on cell viability.

Beas2B cells were grown in 50% DMEM (Life Technologies) and 50% F-12(Life Technologies) media, supplemented with 10% Fetal Bovine Serum(ATCC), 25 mM HEPES (Life Technologies), and 1× Pen-Strep (LifeTechnologies). Cells were cultured in a humidified incubator set at 37°C., 5% CO₂, and trypsinized using 0.25% Trypsin with 0.5%polyvinylpyrrolidone (PVP).

For the assay, Beas2B cells were seeded at 500 cells/well (25 μL/well)in a 384-well plate and cultured overnight. Compounds were seriallydiluted in DMSO, then further diluted with growth media (40 μL/well) tocreate a compound plate 6× of the final assay concentration, at 0.6%DMSO. The diluted compounds were then added to the cells (5 μL/well) andincubated at 37° C., 5% CO₂ for 48 hours. After the compound incubation,CellTiter-Glo (Promega) was added directly to the cells (30 μL/mL). Theassay plate was sealed and shaken at 700 rpm for 15 minutes in adarkened environment, then centrifuged for 2 minutes at 1500 rpm tosettle the lysate at the bottom of the well. The effect of the compoundon cell viability was determined through analysis of dose-dependentquantified changes in ATP from baseline (non-compound treated cells) andwells treated with 60 μM AT9283, a well-characterized cytotoxiccompound. Data are expressed as pCC₁₅ (negative decadic logarithm CC₁₅)values. Certain compounds disclosed herein exhibited pCC₁₅ values ofless than 6 or less than 5.5 when tested in Beas2B cells.

Table 5 shows cytotoxicities of selected compounds in a prematurechromosome condensation assay. Compound numbers correspond to thenumbers and structures provided in Tables 1 and 2 and Examples 1-44.

TABLE 5 ≤5(+++) 5.1 to 5.7 (++) 5.8 to 6.4 (+) Cytotoxicity 5, 6, 10,12, 14, 17, 19, 22, 26, 29, 30, 1, 2, 3, 4, 7, 8, 9, 11, 15, 18, 13, 16,35, 46, pCC₁₅ 31, 33, 36, 37, 40, 42, 43, 44, 47, 48, 20, 21, 23, 24,25, 27, 28, 90, 99, 109, 50, 52, 53, 54, 56, 57, 58, 59, 60, 62, 32, 34,38, 39, 41, 45, 49, 123, 137, 143, 63, 67, 69, 71, 73, 76, 78, 82, 83,84, 51, 55, 61, 64, 65, 66, 68, 151, 153, 156, 86, 87, 88, 89, 92, 93,95, 97, 98, 100, 70, 72, 74, 75, 77, 79, 80, 160, 179, 197, 101, 103,105, 106, 107, 108, 114, 81, 85, 91, 94, 96, 102, 104, 223, 231, 246,118, 119, 121, 122, 124, 125, 126, 110, 111, 112, 113, 115, 252, 253,255, 128, 130, 134, 135, 136, 138, 139, 116, 117, 120, 127, 129, 314,326, 343, 140, 141, 142, 146, 148, 149, 150, 131, 132, 133, 144, 145,351, 379, 393, 152, 154, 155, 157, 159, 161, 162, 147, 158, 165, 166,175, 398, 412, 415, 163, 164, 167, 168, 169, 170, 171, 177, 183, 187,188, 189, 418, 436, 445, 172, 173, 174, 176, 178, 180, 181, 190, 191,192, 193, 194, 446, 447, 450, 182, 184, 185, 186, 195, 200, 201, 196,198, 199, 202, 204, 466, 470, 493, 203, 206, 208, 209, 210, 211, 212,205, 207, 213, 216, 217, 496, 515, 556, 214, 215, 220, 222, 224, 225,226, 218, 219, 221, 233, 234, 1039 227, 228, 229, 230, 232, 235, 236,239, 240, 241, 242, 243, 237, 238, 244, 245, 248, 250, 254, 247, 249,251, 257, 259, 256, 258, 261, 264, 265, 267, 268, 260, 262, 263, 266,270, 269, 271, 272, 273, 275, 276, 281, 274, 277, 278, 279, 280, 282,283, 284, 287, 288, 290, 292, 285, 286, 289, 291, 293, 294, 295, 296,297, 298, 299, 303, 300, 301, 302, 304, 305, 306, 307, 308, 309, 310,311, 313, 312, 315, 317, 319, 321, 316, 318, 320, 323, 325, 327, 328,322, 324, 331, 332, 333, 329, 330, 336, 337, 338, 339, 340, 334, 335,342, 344, 345, 341, 348, 349, 353, 354, 357, 358, 346, 347, 350, 352,355, 363, 364, 365, 368, 369, 371, 373, 356, 359, 360, 361, 362, 378,380, 383, 385, 386, 387, 388, 366, 367, 370, 372, 374, 390, 391, 392,400, 401, 402, 404, 375, 376, 377, 381, 382, 406, 407, 410, 411, 413,416, 419, 384, 389, 394, 395, 396, 420, 421, 422, 423, 425, 426, 427,397, 399, 403, 405, 408, 428, 430, 431, 433, 435, 437, 439, 409, 414,417, 424, 429, 440, 441, 443, 444, 448, 452, 454, 432, 434, 438, 442,449, 455, 456, 458, 460, 465, 467, 468, 451, 453, 457, 459, 461, 472,473, 475, 477, 483, 484, 485, 462, 463, 464, 469, 471, 486, 488, 490,491, 495, 497, 498, 474, 476, 478, 479, 480, 502, 505, 507, 508, 510,511, 512, 481, 482, 487, 489, 492, 513, 517, 518, 520, 527, 530, 532,494, 499, 500, 501, 503, 535, 536, 537, 538, 541, 545, 546, 504, 506,509, 514, 516, 547, 548, 549, 550, 551, 552, 555, 519, 521, 522, 523,524, 557, 558, 560, 561, 562, 564, 565, 525, 526, 528, 529, 531, 567,569, 570, 573, 574, 575, 576, 533, 534, 539, 540, 542, 577, 578, 579,580, 581, 582, 584, 543, 544, 553, 554, 563, 585, 587, 589, 592, 593,594, 595, 566, 568, 571, 572, 583, 596, 597, 601, 602, 603, 605, 606,586, 588, 590, 591, 598, 608, 610, 611, 612, 613, 614, 615, 599, 600,604, 607, 609, 616, 617, 618, 620, 622, 624, 625, 619, 621, 623, 626,627, 628, 629, 630, 632, 633, 634, 636, 631, 635, 637, 638, 639, 641,642, 1000, 1001, 1002, 1007, 640, 1003, 1004, 1005, 1008, 1010, 1011,1012, 1017, 1018, 1006, 1009, 1013, 1014, 1019, 1020, 1021, 1022, 1023,1024, 1015, 1016, 1026, 1027, 1025, 1029, 1034, 1035, 1036, 1043, 1028,1030, 1031, 1032, 1044, 1045 1033, 1037, 1038, 1040, 1041, 1042

Example 48: In Vitro Human Liver Microsome Intrinsic Clearance (HLMCl_(int))

Liver microsomes were used for in vitro determination of hepaticclearance of compounds of the present disclosure. A microsomalincubation cofactor solution was prepared with 100 mM potassiumphosphate buffered to pH 7.4 (BD Biosciences, Woburn, MA) supplementedwith 2 mM NADPH (Sigma-Aldrich, St. Louis, MO). 10 mM DMSO stocks oftest compound were diluted and spiked into the cofactor solution toyield a 0.2 μM concentration (0.02% v/v DMSO). Aliquots of frozen humanliver microsomes (Bioreclamation IVT, Baltimore MD) were thawed anddiluted into 100 mM potassium phosphate buffer to yield microsomalprotein concentrations of 0.2 mg/mL. Cofactor/drug and microsomalsolutions were pre-warmed separately for 4 minutes in a water bath heldat 37° C. Incubations (n=1) were started by the combination of equalvolumes of cofactor/drug solution with microsomal solution. The finalconcentration of test compound was 0.1 μM with a final proteinconcentration of 0.1 mg/mL and final NADPH concentration of 1 mM.Samples were collected at times 0, 3, 8, 15, 30, and 45 minutes tomonitor the disappearance of test compound. At each time point, 50 μL ofincubation sample was removed and spiked into 25 μL of water plus 3%formic acid plus Internal Standard for reaction termination. Sampleswere then injected onto an AB Sciex API 4000 triple quadrupole massspectrometer for quantitation by LC-MS/MS. Mobile Phase A consisted ofHPLC grade water with 0.2% formic acid and Mobile Phase B consisted ofHPLC grade acetonitrile with 0.2% formic acid with all samples runthrough a Thermo HyPURITY C18 50×2.1 mm column (Waltham, MA). HLMCl_(int) data was reported in units of μL/min/mg. See Riley, R. J., etal., Drug Metab. Dispos., 2005, September, 33(9), pp. 1304-1311. Certaincompounds disclosed herein exhibited HLM Cl_(int) of greater than 50μL/min/mg or greater than 100 μL/min/mg.

Example 49: Lung PK/PD

In-Life Portion

C57bl/6n mice were acclimated for at least 3 days before use. On the dayof the experiment, animals were grouped into sample sizes of 5 (n=10 forthe TGF-β stimulated group). Compounds of the present disclosure(formulated in 3% glycerol in PBS; pH=4) were pre-treated via oralaspiration (OA; animals are forced to aspirate solution into the lungsby covering their nose). All oral aspirations were performed using a 50μL dosing volume and accompanied by the appropriate vehicle controlgroups. Following compound OA treatment, the animals were returned totheir home cages and monitored. Compound pre-treatment occurred 4 hoursprior to harvest for screening and dose-response studies; durationstudies had variable compound pre-treatment times. One hour prior toharvest, animals were challenged via oral aspiration a second time withPBS vehicle or recombinant human TGF-β1 protein (0.01 μg per animaldissolved in 1 part 4 mM HCl and 2 parts 3% glycerol in PBS). Fiveminutes prior to harvest, animals were deeply anesthetized underisoflurane and euthanized via cervical dislocation. Bronchoalveolarlavage fluid (BALF), plasma and left lung lobes were collected duringharvest.

Sample Collection and Processing

Blood plasma was collected via open cardiac puncture. After whole bloodcollection, the samples were placed in EDTA-coated tubes to preventcoagulation. Blood samples were spun at 15300×g's for 4 minutes at 4° C.to separate the plasma. Plasma was immediately isolated, frozen andsubmitted for bioanalytical (BA) analysis.

In order to collect BALF, the lungs were flushed via the trachea with0.7 mL of PBS 3 times. The BALF, which consists almost entirely oftissue-derived macrophages, was immediately centrifuged at 700×g's for15 minutes. After centrifugation, the supernatant was removed, the BALFwas re-suspended in 1×cell lysis buffer, and immediately frozen. Priorto BA submission, the BALF was dethawed and sonicated for 30 minutes oncold water to lyse open the cells

Left lung lobes were harvested immediately after BALF collection. Lungsamples were homogenized in 500 μL of 1×cell lysis buffer. Afterhomogenization, the samples were split: half of the sample wasimmediately placed on a rotisserie for 10 minutes while the other halfwas immediately frozen for BA analysis. The samples placed on therotisserie were then centrifuged at 10,000×g's for 10 minutes in orderto separate the protein in the supernatant from pelleted debris.Following collection of the supernatant, a total protein quantificationassay (Bradford) was performed to normalize the concentrations of allsamples. Using the Hamilton star liquid handling system, each sample wasdiluted in 1×cell lysis buffer to 2 mg/mL of protein. Samples werestored at −80° C. or immediately processed using the Meso-scaleDiscovery system.

Phospho-SMAD3 (pSMAD3) and Total-SMAD3 (tSMAD3) Quantification usingMeso-Scale Discovery

Meso-scale Discovery (MSD) is an electrochemical protein quantificationassay that requires specialized microplates with carbon electrodesattached to the bottom. These carbon electrodes allow for greaterattachment of biological reagent to microplates, thus allowing for amore sensitive read-out when compared to a traditional ELISA. Similar toa standard sandwich ELISA, MSD requires use of a coating antibody thatbinds the target protein(s) within the sample. After sample incubation,a primary antibody is used to bind the epitope of interest. Followingaddition of the primary antibody, a secondary-antibody with a SULFO-TAGdetection is used to allow for quantification of the epitope ofinterest. Lastly, the microplate is read via an electric pulse thatcauses the SULFO-TAG to emit light, which serves as the final read-outof the assay.

The coating antibody (SMAD3, clone=5G-11) was incubated overnight in thespecialized MSD microplates at 4° C. The next day, the microplates wereblocked in 3% BSA (bovine serum albumin) for 70 minutes to preventnon-specific protein binding to the bottom of the microplate. After awash step, 50 μg of lung samples were loaded into the MSD-plate andincubated for 2 hours at room temperature. The plates were washed againto remove unbound sample; either phospo-SMAD3 (pSMAD3; clone=EP568Y) ortotal-SMAD3 (tSMAD3) primary antibody were incubated for 1 hour.Following a wash step, the anti-rabbit SULFO-tag detection antibody wasincubated for 50 minutes. After a final wash step, MSD-read buffer wasadded to each sample. pSMAD3 and tSMAD3 quantification was performedusing an MSD-specific plate reader (Sector S 600).

Data Analysis

Samples were immediately analyzed using an outlier analysis (Grubbstest, α=0.05). After outlier removal, the raw pSMAD3 were divided by thetSMAD3 luminescent readings. In screening and dose-response studies, thepSMAD3/tSMAD3 ratio was normalized to the TGF-β induction group (set to100%) in order to minimize the variability between stimulation. First,the 3% glycerol/PBS group was compared with the 3% glycerol/TGF-β with astudent's t-test (cut-off: p=0.05) to ensure a pSMAD3 window waspresent. A one-way ANOVA (fisher's uncorrected LSD) was used to compareall drug treated groups with the 3% glycerol/TGF-β group to determine ifstatistically significant differences are observed. Percent pSMAD3inhibition was calculated using the vehicle pSMAD3 as a baseline valueand displayed as the final readout. Dose-response curves were fittedwith a 4-parameter non-linear regression algorithm; the minimum responsewas set to 0% pSMAD3 inhibition and the maximum response set to 100%pSMAD3 inhibition. Compound potencies were obtained from the regressionand reported as ID50s.

PK Study

Plasma, lung and macrophage drug concentrations were quantified. Totalmacrophage concentration was normalized to the total macrophage cellvolume over the total drug recovered in the BALF. The alveolarmacrophage volume used in the calculation was based on a publication byKrombach et al. (Environmental Health Perspectives, September 1997, Vol.105, Supplement 5, pp. 1261-1263) which estimated the rat alveolarmacrophage volume to be approximately 1200 μm³ or 1.2e⁻⁹ mL. Theassumption was made that the mouse alveolar macrophage volume is similarto that of the rat. Normalized total macrophage concentrationrecovered=(total drug recovered from BALF)/(total cell counts*1.2e⁻⁹mL).

Certain compounds disclosed herein exhibited (lung AUC_(0-t)):(plasmaAUC_(0-t)) ratios of greater than 10, such as greater than 50, greaterthan 75 or greater than 100. A compound intended for local delivery tothe lung with minimal systemic exposure preferably exhibits a (lungAUC_(0-t)):(plasma AUC_(0-t)) ratio of greater than 50. Certaincompounds provided in Table 3 having pK_(i) values of greater than 9.5exhibited a (lung AUC_(0-t)):(plasma AUC_(0-t)) ratio of greater than75.

Example 50: Cardiac PK/PD

In-Life Portion

C57bl/6n mice were acclimated for at least 3 days before use. On the dayof the experiment, animals were grouped into sample sizes of 5-10. Testcompounds were pre-treated via oral aspiration (OA; animals are forcedto aspirate solution into the lungs by covering their nose). All oralaspirations were performed using a 50 μL dosing volume and accompaniedby a vehicle control group (3% glycerol in PBS, pH=4). Followingcompound OA treatment, the animals were returned to their home cages andmonitored. Compound pre-treatment occurred either 2 or 4 hours prior toharvest. One hour prior to harvest, animals were challenged viatail-vein intravenous injection with PBS vehicle or recombinant humanTGF-β1 protein (1 μg per animal dissolved in 1 part 4 mM HCl and 2 parts3% glycerol in PBS). Five minutes prior to harvest, animals were deeplyanesthetized under isoflurane and euthanized via cervical dislocation.Plasma, left lung lobes and whole hearts were collected during harvest.

Sample Collection and Processing

Blood plasma was harvested as described above in the Lung PK/PDexperiment. Whole hearts were processed in the same manners as left lunglobes in the Lung PK/PD experiment. Left lung lobes were homogenized in500 μL of water and submitted for BA Analysis.

Phospho-SMAD3 (pSMAD3) and Total-SMAD3 (tSMAD3) Quantification UsingMeso-Scale Discovery

Heart samples were processed using MSD in the same manner as the leftlung lobes above. Data analysis was performed in the same manners as thelung PK/PD experiment. Plasma, lung and heart drug concentrations werequantified.

There was minimal target engagement systemically following treatmentwith one or more compound disclosed herein, as measured by SMAD3phosphorylation inhibition. In some examples, a compound disclosedherein exhibited less than 10% target engagement systemically asmeasured by SMAD3 phosphorylation inhibition.

Example 51: Efficacy Study in Syngeneic Cancer Model

One or more compounds disclosed herein, e.g., a compound provided inTable 1 having an ALK5 pK_(i) value of greater than 9.5, preferablygreater than 10.5 (a measurement reflecting the ability of the compoundto inhibit ALK5 activity, measured in accordance with Example 45), areexpected to suppress tumor growth in syngeneic cancer models whenadministered alone or in combination with an immunotherapeutic agent.Six- to 8-week old BALB/c mice are used for in vivo efficacy studies inaccordance with IACUC guidelines. Commercially available 4T1 cells(0.5-2.0×10⁴ cells/mouse) are implanted subcutaneously into the rightflanks of BALB/c mice. When the tumor reaches a palpable size ofapproximately 8-10 mm in diameter, the primary tumors are surgicallyremoved, and the mice are randomly assigned to vehicle control orcompound treatment groups. Alternatively, CT26 cells (0.5-2.0×10⁴cells/mouse) are injected intravenously into BALB/c mice to generate thecancer model. Two days following the surgery, or 7 days followinginjection of CT26 cells, the mice are treated with either (1) vehiclecontrol, (2) a compound of the present disclosure at an appropriateamount and frequency (formulated in 3% glycerol in PBS; pH=4) via oralaspiration or intranasally, (3) an immunotherapeutic agent (e.g.,pembrolizumab or durvalumab) at an appropriate amount and frequency, or(4) a compound of the present disclosure and an immunotherapeutic agent,each at an appropriate amount and frequency.

Body weight is measured twice weekly. Following 2- to 4-weeks oftreatment, the lung and liver of each animal is harvested, and thenumber of metastatic cells in each tissue sample determined using aclonogenic metastasis assay. Cells may be further subjected to one ormore of FACS analysis, T-cell function assay, and RNA extraction. It isexpected that the animal group treated with one or more of the ALK5inhibitors disclosed herein exhibits reduction in lung tumor burden.Activation of an immune response by the ALK5 inhibitor may stimulateboth local and systemic antitumor T-cell activation, thus a reduction inliver tumor burden may also be observed. When administered incombination with an immunotherapeutic agent, a compound of the presentdisclosure, such as a compound provided in Table 1, is expected toproduce an increased reduction in lung tumor burden relative to thereduction in tumor burden observed in animals treated with either singleagent alone. The compounds described herein are expected to interactsynergistically with an immunotherapeutic agent to suppress tumor growthand increase survival.

Example 52: Prophylactic Study in Murine DSS-Induced Intestinal FibrosisModel

One or more compounds disclosed herein, e.g., a compound provided inTable 1 having an ALK5 pK_(i) value of greater than 9.5, preferablygreater than 10.5 (a measurement reflecting the ability of the compoundto inhibit ALK5 activity, measured in accordance with Example 45), areexpected to slow, halt or reverse the progression of intestinal fibrosisin a murine colitis model. Six to 8-week old male C57BL/6J mice aretagged and weighed. The drinking water of the animals is treated with2.5% dextran sulfate sodium (DSS) for 7 days to induce acute colitis,followed by 2 days of normal drinking water. Three, 3-week cycles of2.5% DSS treatment (1 week of 2.5% DSS in water; 2 weeks of normalwater) are then completed to induce intestinal fibrosis.

Starting on day one of DSS administration, mice are treated with eithervehicle control or a compound of the present disclosure at anappropriate amount and frequency via oral gavage (e.g., once daily). Theanimals are sacrificed 9 weeks after the first DSS administration, thendistal, mid and proximal sections of the colon harvested for histologicanalysis, RNA extraction and cytokine measurement. A compound of thepresent disclosure, such as a compound provided in Table 1, is expectedto decrease ALK5 activity in the colon and to slow or prevent intestinalfibrosis as evidenced by one or more of (1) reduction in the ratio ofcolon weight to colon length; (2) reduction in deposition ofextracellular matrix as observed by histology; (3) reduction inexpression of collagen 1 (Colla1) and connective tissue growth factor(Ctgf) in colon tissue; and (4) reduction in production of TGF-β1 andIL6 in the colon, relative to vehicle-treated controls.

Example 53: Efficacy Study in Murine DSS-Induced Intestinal FibrosisModel

One or more compounds disclosed herein, e.g., a compound provided inTable 1 having an ALK5 pK_(i) value of greater than 9.5, preferablygreater than 10.5 (a measurement reflecting the ability of the compoundto inhibit ALK5 activity, measured in accordance with Example 45), areexpected to slow, halt or reverse the progression of intestinal fibrosisin a murine colitis model. Six to 8-week old male C57BL/6J mice aretagged and weighed. The drinking water of the animals is treated with2.5% dextran sulfate sodium (DSS) for 7 days to induce acute colitis,followed by 2 days of normal drinking water. Three, 3-week cycles of2.5% DSS treatment (1 week of 2.5% DSS in water; 2 weeks of normalwater) are then completed to induce intestinal fibrosis.

Following the second of the 3 cycles of DSS administration, mice aretreated with either vehicle control or a compound of the presentdisclosure at an appropriate amount and frequency via oral gavage (e.g.,once daily). Animals are sacrificed at either 6, 9 or 12 weeks after thefirst DSS cycle, then distal, mid and proximal sections of the colonharvested for histologic analysis, RNA extraction and cytokinemeasurement. A compound of the present disclosure, such as a compoundprovided in Table 1, is expected to decrease ALK5 activity in the colonand to slow, halt or reverse intestinal fibrosis as evidenced by one ormore of (1) reduction in the ratio of colon weight to colon length; (2)reduction in deposition of extracellular matrix as observed byhistology; (3) reduction in expression of collagen 1 (Col1a1) andconnective tissue growth factor (Ctgf) in colon tissue; and (4)reduction in production of TGF-β1 and IL6 in the colon, relative tovehicle-treated controls.

Example 54: Efficacy Study in Adoptive T-Cell Transfer Model of Colitis

One or more compounds disclosed herein, e.g., a compound provided inTable 1 having an ALK5 pK_(i) value of greater than 9.5, preferablygreater than 10.5 (a measurement reflecting the ability of the compoundto inhibit ALK5 activity, measured in accordance with Example 45), areexpected to slow, halt or reverse the progression of intestinal fibrosisin an adoptive T-cell transfer model of colitis. Six- to 8-week oldfemale CB17 SCID mice are tagged and weighed, then administered CD4⁺CD25⁻ CD62L⁺ naïve T cells isolated from the spleens of Balb/C mice (IP;1×10⁶ cells) to induce colitis.

Once diarrhea and a 10% or greater decrease in body weight are observed(typically around week 2), mice are treated with either vehicle controlor a compound of the present disclosure at an appropriate amount andfrequency via oral gavage (e.g., once daily). Animals are sacrificed 45days after induction of colitis, then distal, mid and proximal sectionsof the colon harvested for histologic analysis, RNA extraction andcytokine measurement. A compound of the present disclosure, such as acompound provided in Table 1, is expected to decrease ALK5 activity inthe colon and to slow, halt or reverse intestinal fibrosis as evidencedby one or more of (1) reduction in the ratio of colon weight to colonlength; (2) reduction in deposition of extracellular matrix as observedby histology; (3) reduction in expression of collagen 1 (Col1a1) andconnective tissue growth factor (Ctgf) in colon tissue; and (4)reduction in production of TGF-β1 and IL6 in the colon, relative tovehicle-treated controls.

Example 55: Efficacy Study in Monocrotaline Model of Severe PulmonaryHypertension

One or more compounds disclosed herein, e.g., a compound provided inTable 1 having an ALK5 pK_(i) value of greater than 9.5, preferablygreater than 10.5 (a measurement reflecting the ability of the compoundto inhibit ALK5 activity, measured in accordance with Example 45), areexpected to slow, halt or reverse the progression of pulmonaryhypertension in a monocrotaline (MCT) model of severe pulmonaryhypertension. Male Sprague-Dawley rats are tagged, weighed, and randomlydivided into control and MCT-treated groups. The rats in the MCT-treatedgroup are administered a single dose of MCT (60 mg/kg, s.c.), thentreated with either (1) vehicle control; (2) sildenafil (30 mg/kg, p.o.,b.i.d.); or (3) a compound of the present disclosure at an appropriateamount and frequency (formulated in 3% glycerol in PBS; pH=4) via oralaspiration.

Following 2-weeks of treatment, the animals are anesthetized withketamine/xylazine for terminal monitoring of pulmonary and systemicarterial pressures along with heart rate. The lungs of each animal arethen harvested for histologic analysis. A compound of the presentdisclosure, such as a compound provided in Table 1, is expected todecrease ALK5 activity in the lung and slow, halt or reverse theprogression of pulmonary hypertension as evidenced by one or more of (1)reduction in systolic pulmonary arterial pressure; (2) reduction inright ventricular (RV) systolic pressure; (3) reduction in RV diastolicpressure; (4) increase in cardiac output; (5) reduction in RVhypertrophy; (6) reduction in pSmad2 or pSmad3 staining within vascularand/or alveolar cells; (7) reduction in medial thickness; (8) reductionin vascular smooth muscle cell proliferation; (9) reduction in vascularsmooth muscle hypertrophy; and (10) reduction in expression of matrixmetalloproteinase (MMP)-2 and/or MMP-9.

1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: A is selected from phenyl, 5- to 6-membered heteroaryl, indanyl, indazolyl, benzotriazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzodioxanyl, and tetrahydrobenzazepinyl; B is selected from phenyl and 5- to 6-membered heteroaryl; R^(A) is independently selected at each occurrence from: halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —S(═O)₂NR²R³, —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂, —C(O)NR²R³; C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from —OR¹, —N(R¹)₂, —C(O)OR¹, C₃₋₈ carbocycle, and 3- to 10-membered heterocycle; and C₃₋₈ carbocycle and 3- to 10-membered heterocycle, wherein each C₃₋₈ carbocycle and 3- to 10-membered heterocycle in R^(A) is independently optionally substituted with one or more substituents selected from —OR¹, —CH₂N(R¹)₂, —N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —C(O)N(R¹)₂, R¹, and C₁₋₆ alkyl; R^(B) is independently selected at each occurrence from halogen, —CN, —NH₂, —NHCH₃, —NHCH₂CH₃, —C(O)CH₃, —OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂F, —CHF₂, —CF₃, C₃₋₄ carbocycle, and 3- to 4-membered heterocycle; m is 0, 1 or 2; n is 0, 1, 2, or 3; R¹ is independently selected at each occurrence from hydrogen; and C₁₋₆ alkyl, 1- to 6-membered heteroalkyl, C₀₋₃ alkyl-(C₃₋₈ carbocycle), and C₀₋₃ alkyl-(3- to 10-membered heterocycle), each of which is optionally substituted by one or more substituents selected from —NH₂, —NHCH₃, —N(CH₃)₂, —CH₂CH₂N(CH₃)₂, —C(O)CH₃, —C(O)OH, —C(O)NH₂, ═O, —OH, —CH₂OH, —CH₂CH₂OH, —OCH₃, —OCH₂CH₃, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, C₃₋₈ carbocycle, and 3- to 6-membered heterocycle; and R² and R³ are taken together with the nitrogen atom to which they are attached to form a heterocycle, optionally substituted with one or more R¹. 2.-3. (canceled)
 4. The compound or salt of claim 1, wherein A is selected from phenyl and 5- to 6-membered heteroaryl.
 5. The compound or salt of claim 1, wherein A is selected from phenyl, indanyl, thiazolyl, thiophenyl, pyrazolyl, pyridyl, pyrimidinyl, indazolyl, benzotriazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzodioxanyl, and tetrahydrobenzazepinyl.
 6. The compound or salt of claim 1, wherein A is phenyl.
 7. The compound or salt of claim 1, wherein the compound is a compound of Formula (I-B):


8. The compound or salt of claim 1, wherein the compound is a compound of Formula (I-C):

9.-10. (canceled)
 11. The compound or salt of claim 1, wherein R^(A) is independently selected at each occurrence from: halogen, —OR¹, —N(R¹)₂, —S(═O)₂R¹, —S(═O)₂N(R¹)₂, —NR¹S(═O)₂R¹, —NR¹S(═O)₂N(R¹)₂, —C(O)R¹, —C(O)OR¹, —NR¹C(O)R¹, —NR¹C(O)N(R¹)₂, —C(O)N(R¹)₂; C₁₋₆ alkyl and —N(R¹)—C₁₋₁₀ alkyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from —OR¹, —N(R¹)₂, and 3- to 10-membered heterocycle; and 3- to 10-membered heterocycle, wherein each 3- to 10-membered heterocycle in R^(A) is independently optionally substituted with one or more substituents selected from —OR¹, —N(R¹)₂, —C(O)R¹, —CH₂C(O)OR¹, —C(O)OR¹, —C(O)N(R¹)₂, and R¹. 12.-13. (canceled)
 14. The compound or salt of claim 1, wherein R^(A) is —NR¹C(O)R¹.
 15. The compound or salt of claim 1, wherein R^(A) is —C(O)OR¹.
 16. The compound or salt of claim 1, wherein R^(A) is C₁₋₆ alkyl substituted with 4- to 8-membered heterocycle, wherein the 4- to 8-membered heterocycle is substituted with —C(O)OR¹. 17.-21. (canceled)
 22. The compound or salt of claim 1, wherein m is
 1. 23. (canceled)
 24. The compound or salt of claim 1, wherein B is selected from phenyl and pyridyl.
 25. (canceled)
 26. The compound or salt of claim 1, wherein B is 2-pyridyl.
 27. (canceled)
 28. The compound or salt of claim 1, wherein R^(B) is independently selected at each occurrence from halogen, —OH, —CH₃, and —CHF₂.
 29. The compound or salt of claim 1, wherein R^(B) is —CH₃.
 30. (canceled)
 31. The compound or salt of claim 1, wherein n is
 1. 32. (canceled)
 33. The compound or salt of claim 1, wherein the compound is a compound of Formula (I-E):

34.-40. (canceled)
 41. A pharmaceutical composition comprising the compound or salt of claim 1 a pharmaceutically acceptable carrier. 42.-43. (canceled)
 44. A method of treating an ALK5-mediated disease or condition in a subject, comprising administering to the subject a therapeutically effective amount of the compound or salt of claim
 1. 45. (canceled)
 46. The method of claim 44, wherein the disease or condition is fibrosis. 47.-50. (canceled)
 51. The method of claim 46, where the fibrosis is idiopathic pulmonary fibrosis (IPF).
 52. The method of claim 46, wherein the fibrosis is intestinal fibrosis.
 53. (canceled)
 54. The method of claim 44, wherein the disease or condition is selected from breast cancer, colon cancer, prostate cancer, lung cancer, hepatocellular carcinoma, glioblastoma, melanoma, and pancreatic cancer. 55.-63. (canceled) 